46 results on '"Sahr, T"'
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2. Legionella pneumophila effector RomA uniquely modifies host chromatin to repress gene expression and promote intracellular bacterial replication
- Author
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Rolando M, Sanulli S, Rusniok C, Gomez-Valero L, Bertholet C, Sahr T, Margueron R, and Buchrieser C.
- Published
- 2013
3. Precision electroweak measurements on the Z resonance
- Author
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Schael, S, Barate, R, Bruneliere, R, Buskulic, D, Bonis, De, Decamp, I, Ghez, D, Goy, P, Jezequel, C, Lees, S, Lucotte, Jp, Martin, A, Merle, F, Minard, E, Nief, Mn, Odier, Jy, Pietrzyk, P, Trocme, B, Bravo, B, Casado, S, Chmeissani, Mp, Comas, M, Crespo, P, Fernandez, Jm, E, Fernandez, Bosman, Garrido, M, Grauges, L, Juste, E, Martinez, A, Merino, M, Miquel, G, Mir, R, Orteu, Lm, Pacheco, S, Park, A, Perlas, Ic, Riu, J, Ruiz, I, Sanchez, H, Colaleo, F, Creanza, A, D, Filippis, De, N, Palma, De, Iaselli, M, Maggi, G, Nuzzo, M, Ranieri, S, Raso, A, Ruggieri, G, Selvaggi, F, Silvestris, G, Tempesta, L, Tricomi, P, Zito, A, Huang, G, Lin, X, Ouyang, J, Wang, Q, Xie, T, Xu, Y, Xue, R, Zhang, S, Zhang, J, Zhao, L, Abbaneo, W, Bazarko, D, Becker, A, Boix, U, Bird, G, Blucher, F, Bonvicini, E, B, Bright, Thomas, Barklow, P, Buchmuller, T, Cattaneo, O, Cerutti, M, Ciulli, F, Clerbaux, V, Drevermann, B, Forty, H, Frank, Rw, Greening, M, Hagelberg, Tc, Halley, R, Gianotti, Aw, Girone, F, Hansen, M, Harvey, Jb, Jacobsen, J, Hutchcroft, R, Janot, De, Jost, R, Knobloch, B, Kado, J, Lehraus, M, Lazeyras, I, Maley, P, Mato, R, May, P, Moutussi, J, A, Pepe, Altarelli, Ranjard, M, Rolandi, F, Schlatter, L, Schmitt, D, Schneider, B, Tejessy, O, Teubert, W, Tomalin, F, Tournefier, Ir, Veenhof, E, Valassi, R, Wiedenmann, A, Wright, W, Ajaltouni, Ae, Badaud, Z, Chazelle, F, Deschamps, G, Dessagne, O, Falvard, S, Ferdi, A, Fayolle, C, Gay, D, Guicheney, P, Henrard, C, Jousset, P, Michel, J, Monteil, B, Montret, S, Pallin, Jc, Pascolo, D, Perret, Jm, Podlyski, P, Bertelsen, F, Fernley, H, Hansen, T, Hansen, Jd, Hansen, Jr, Kraan, Ph, Lindahl, Ac, Mollerud, A, Nilsson, R, Rensch, Bs, Waananen, B, Daskalakis, A, Kyriakis, G, Markou, A, Simopoulou, C, Siotis, E, Vayaki, I, Zachariadou, A, Blondel, K, Bonneaud, A, Brient, G, Machefert, Jc, Rouge, E, Rumpf, A, Swynghedauw, M, Tanaka, M, Verderi, R, Videau, M, Focardi, H, Parrini, E, Corden, G, Georgiopoulos, M, Antonelli, C, Antonelli, A, Bencivenni, M, Bologna, G, Bossi, G, Campana, F, Capon, P, Chiarella, G, Felici, V, Laurelli, G, Mannocchi, P, Murtas, G, Passalacqua, Gp, Picchi, L, Colrain, P, P, Ten, Have, Hughes, I, Kennedy, Is, Knowles, J, Lynch, Ig, Morton, Jg, Negus, Wt, Oshea, P., Raine, V, Reeves, C, Scarr, P, Smith, Jm, Thompson, K., Turnbull, As, Wasserbaech, Rm, Cavanaugh, S, Dhamotharan, R, Geweniger, S, Hanke, C, Hansper, P, Hepp, G, Kluge, V, Putzer, Ee, Sommer, A, Stenzel, J, Tittel, H, Werner, K, Wunsch, W, Beuselinck, M, Binnie, R, Cameron, Dm, Davies, W, Dornan, G, Goodsir, Pj, Marinelli, S, Martin, N, Nash, Eb, Nowell, J, Rutherford, J, Sedgbeer, Sa, Thompson, Jk, White, Jc, Williams, R, Ghete, Md, Girtler, Vm, Kneringer, P, Kuhn, E, Rudolph, D, G, Bouhova, Thacker, Bowdery, E, Buck, Ck, Clarke, Pg, Ellis, Dp, Finch, G, Foster, Aj, Hughes, F, Jones, G, Rwl, Keemer, Pearson, Nr, Robertson, Mr, Sloan, Na, Smizanska, T, Snow, M, Williams, Sw, van der Aa, Delaere, O, Leibenguth, C, Lemaitre, G, Bauerdick, V, Lat, Blumenschein, U, Van, Gemmeren, Giehl, P, Holldorfer, I, Jakobs, F, Kasemann, K, Kayser, M, Kleinknecht, F, Muller, K, Quast, As, Renk, G, Rohne, B, Sander, E, Schmeling, Hg, Wachsmuth, S, Wanke, H, Zeitnitz, R, Ziegler, C, Aubert, T, Benchouk, Jj, Bonissent, C, Carr, A, Coyle, J, Curtil, P, Ealet, C, Etienne, A, Fouchez, F, Motsch, D, Payre, F, Rousseau, P, Tilquin, D, Talby, A, Thulasides, M, Aleppo, M, Ragusa, M, Buscher, F, David, V, Dietl, A, Ganis, H, Huttmann, G, Lutjens, K, Mannert, G, Manner, C, Moser, W, Settles, Hg, Seywerd, R, Villegas, H, Wolf, M, Azzurri, G, Boucrot, P, Callot, J, Chen, O, Cordier, S, Davier, A, Duflot, M, Grivaz, L, Heusse, Jf, Jacholkowska, P, Diberder, Le, Lefrancois, F, Mutz, J, Schune, Am, Serin, Mh, Veillet, L, Videau, Jj, Zerwas, I, Bagliesi, D, Bettarini, G, Boccali, S, Bozzi, T, Calderini, C, Dellorso, G, Fantechi, R, Ferrante, R, Fidecaro, I, Foa, F, Giammanco, L, Giassi, A, Gregorio, A, Ligabue, A, Lusiani, F, Marrocchesi, PIER SIMONE, Messineo, Ps, Palla, A, Rizzo, F, Sanguinetti, G, Sciaba, G, Sguazzoni, A, Spagnolo, G, Steinberger, P, Tenchini, J, Venturi, R, Vannini, A, Verdini, C, Awunor, Pg, Blair, O, Cowan, Ga, Garcia, Bellido, Green, A, Medcalf, Mg, Misiejuk, T, Strong, A, Teixeira, Dias, Botterill, P, Clifft, Dr, Edgecock, Rw, Edwards, Tr, Haywood, M, Norton, Sj, Ward, Pr, Bloch, Devaux, Boumediene, B, Colas, D, Emery, P, Fabbro, S, Kozanecki, B, Lancon, W, Lemaire, E, Locci, Mc, Perez, E, Rander, P, Renardy, J, Roussarie, Jf, Schuller, A, Schwindling, Jp, Tuchming, J, Vallage, B, Black, B, Dann, Sn, Kim, Jh, Konstantinidis, Hy, Litke, N, Mcneil, Am, Taylor, Ma, Booth, G, Cartwright, Cn, Combley, S, Hodgson, F, Lehto, Pn, Thompson, M, Affholderbach, Lf, Barberio, K, Bohrer, E, Brandt, A, Burkhardt, S, Feigl, H, Grupen, E, Hess, C, Lutters, J, Meinhard, G, H, Minguet, Rodriguez, Mirabito, J, Neugebauer, L, Ngac, E, Prange, A, Rivera, G, Saraiva, F, Schafer, P, Sieler, U, Smolik, U, Stephan, L, Trier, F, Apollonio, H, Borean, M, Bosisio, C, L, Della, Marina, Giannini, R, Gobbo, G, Musolino, B, Pitis, G, He, L, Kim, H, Putz, H, Rothberg, J, Armstrong, J, Bellantoni, Sr, Berkelman, L, Cinabro, K, Conway, D, Cranmer, Js, Elmer, K, Feng, P, Ferguson, Z, Dps, Gao, Gonzalez, Y, Grahl, S, Harton, J, Hayes, Jl, Hu, Oj, Jin, H, Johnson, S, Kile, Rp, Mcnamara, J, Nielsen, Pa, Orejudos, J, Pan, W, Saadi, Y, Scott, Y, Sharma, Ij, Walsh, V, Walsh, Am, Wear, J, J, von Wimmersperg Toeller, Wu, Jh, Wu, J, Wu, Sl, Yamartino, X, Zobernig, Jm, Dissertori, G, Abdallah, G, Abreu, J, Adam, P, Adye, W, Adzic, T, Ajinenko, P, Albrecht, I, Alderweireld, T, Alekseev, T, Alemany, Fernandez, Allmendinger, R, Allport, T, Almehed, Pp, Amaldi, S, Amapane, U, Amato, N, Anashkin, S, Anassontzis, E, Andersson, Eg, Andreazza, P, Andringa, A, Anjos, S, Antilous, N, Apel, P, Arnoud, Wd, Ask, Y, Asman, S, Augustin, B, Augustinus, Je, Baillon, A, Ballestrero, P, Bambade, A, Barao, P, Barbiellini, F, Barbier, G, Bardin, R, Barker, D, Baroncelli, G, Battaglia, A, Baubillier, M, Becks, M, Begalli, Kh, Behrmann, M, Beilliere, A, Belokopytov, P, Belous, Y, K, Ben, Haim, Benekos, E, Benvenuti, N, Berat, A, Berggren, C, Berntzon, M, Bertini, L, Bertrand, D, Besancon, D, Besson, M, Bianchi, N, Bigi, F, Bilenky, M, Bizouard, Ms, Bloch, Ma, Blom, D, Bluj, M, Bonesini, M, Bonivento, M, Boonekamp, W, Booth, M, Psl, Borgland, Borisov, Aw, Bosio, G, Botner, C, Boudinov, O, Bouquet, E, Bourdarios, B, Bowcock, C, Tjv, Boyko, Bozovic, I, Bozzo, I, Bracko, M, Branchini, M, Brenke, P, Brenner, T, Brodet, R, Bruckman, E, Brunet, P, Bugge, Jm, Buran, L, Burgsmueller, T, Buschbeck, T, Buschmann, B, Cabrera, P, Caccia, S, Calvi, M, Rozas, M, Ajc, Camporesi, Canale, T, Canepa, V, Carena, M, Carroll, F, Caso, L, Gimenez, C, Mvc, Castro, Cattai, N, Cavallo, A, Cerruti, F, Chabaud, C, Chapkin, V, Charpentier, M, Chaussard, P, Checchia, L, Chelkov, P, Chen, Ga, Chierici, M, Chliapnikov, R, Chochula, R, Chorowicz, P, Chudoba, V, Chung, J, Cieslik, Su, Collins, K, Colomer, P, Contri, M, Cortina, R, Cosme, E, Cossuti, G, Costa, F, Cowell, Mj, Crawley, Jh, Crennell, Hb, Crepe, D, Crosetti, S, Cuevas, G, Czellar, J, Dhondt, S, Dalmagne, J, Dalmau, B, Damgaard, J, Davenport, G, M, Silva, Da, T, Deghorain, W, Della, Ricca, Delpierre, G, Demaria, P, Angelis, De, Boer, De, W, Brabandere, De, S, Clercq, De, C, Lotto, De, Maria, De, Min, De, Paula, De, Dijkstra, L, Ciaccio, Di, Diodato, Di, Simone, Di, Djannati, A, Dolbeau, A, Doroba, J, Dracos, K, Drees, M, Drees, J, Dris, Ka, Duperrin, M, Durand, A, Ehret, Jd, Eigen, R, Ekelof, G, Ekspong, T, Ellert, G, Elsing, M, Engel, M, Erzen, Jp, Santo, B, Mce, Falk, Fanourakis, E, Fassouliotis, G, Fayot, D, Feindt, J, Fenyuk, M, Fernandez, A, Ferrari, J, Ferrer, P, Ferrer, Ribas, Ferro, E, Fichet, F, Firestone, S, Fischer, A, Flagmeyer, Pa, Foeth, U, Fokitis, H, Fontanelli, E, Franek, F, Frodesen, B, Fruhwirth, Ag, R, Fulda, Quenzer, Fuster, F, Galloni, J, Gamba, A, Gamblin, D, Gandelman, S, Garcia, M, Garcia, C, Gaspar, J, Gaspar, C, Gasparini, M, Gavillet, U, Gazis, P, Gele, E, Gerber, D, Gerdyukov, Jp, Ghodbane, L, Gil, N, Glege, I, Gokieli, F, Golob, R, Gomez, Ceballos, Goncalves, G, Caballero, P, Gopal, Ig, Gorn, G, Gorski, L, Gouz, M, Gracco, Y, Graziani, V, Green, E, Grefrath, C, Grimm, A, Gris, Hj, Grosdidier, P, Grzelak, G, Gunther, K, Guy, M, Haag, J, Hahn, C, Hahn, F, Hallgren, S, Hamacher, A, Hamilton, K, Hansen, K, Harris, J, Haug, Fj, Hauler, S, Hedberg, F, Heising, V, Hennecke, S, Henriques, M, Hernandez, R, Herquet, Jj, Herr, P, Hessing, H, Heuser, Tl, Higon, Jm, Hoffman, E, Holmgren, J, Holt, So, Holthuizen, Pj, Hoorelbeke, D, Houlden, S, Hrubec, Ma, Huber, J, Huet, M, Hughes, K, Hultqvist, Gj, Jackson, K, Jacobsson, Jn, Jalocha, R, Janik, P, Jarlskog, R, Jarlskog, C, Jarry, G, Jean, Marie, Jeans, B, Johansson, D, Johansson, Ek, Jonsson, Pd, Joram, P, Juillot, C, Jungermann, P, Kapusta, L, Karafasoulis, F, Katsanevas, K, 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Thaler, Fe, Thom, Jj, Trandafir, J, Turk, Ai, Usher, Jd, Vavra, T, Vella, J, Venuti, E, Verdier, Jp, Wagner, R, Waite, Sr, Walston, Ap, Wang, S, Watts, J, Weidemann, Sj, Weiss, Aw, Whitaker, Er, White, Js, Wickens, Sl, Williams, Fj, Williams, Da, Williams, Dc, Willocq, Sh, Wilson, S, Wisniewski, Rj, Wittlin, Wj, Woods, Jl, Word, M, Wright, Gb, Wyss, Tr, Yamamoto, J, Yang, Rk, Yashima, Xq, Yellin, J, Young, Sj, Yuta, Cc, Zapalac, H, Zdarko, G, Zeitlin, Rw, Zhou, C, Blondel, Alain, Schael, S, Barate, R, Bruneliere, R, Spagnolo, Stefania Antonia, S., Schael, Aloisio, Alberto, Alviggi, Mariagrazia, Canale, Vincenzo, Chiefari, Giovanni, DELLA VOLPE, Domenico, Merola, Leonardo, Napolitano, Marco, Patricelli, Sergio, Sciacca, Crisostomo, Lista, Luca, Laboratoire d'Annecy de Physique des Particules (LAPP), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université Savoie Mont Blanc (USMB [Université de Savoie] [Université de Chambéry])-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Physique Corpusculaire - Clermont-Ferrand (LPC), Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Physique Théorique et Astroparticules (LPTA), Université Montpellier 2 - Sciences et Techniques (UM2)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Laboratoire Leprince-Ringuet (LLR), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-École polytechnique (X)-Centre National de la Recherche Scientifique (CNRS), Centre de Physique des Particules de Marseille (CPPM), Aix Marseille Université (AMU)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de l'Accélérateur Linéaire (LAL), Université Paris-Sud - Paris 11 (UP11)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Physique Nucléaire et de Hautes Énergies (LPNHE), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Physique Subatomique et de Cosmologie (LPSC), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Institut Polytechnique de Grenoble - Grenoble Institute of Technology-Centre National de la Recherche Scientifique (CNRS), Institut de Physique Nucléaire de Lyon (IPNL), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Institut de Recherches Subatomiques (IReS), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Cancéropôle du Grand Est-Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), Physique Corpusculaire et Cosmologie - Collège de France (PCC), Collège de France (CdF (institution))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), ALEPH, DELPHI, L3, OPAL, SLD, BARBIELLINI AMIDEI, Guido, Bosisio, Luciano, DELLA RICCA, Giuseppe, Giannini, Gianrossano, Gregorio, Anna, Lanceri, Livio, Poropat, Paolo, Vitale, Lorenzo, Buskulic, D, De Bonis, I, Decamp, D, Ghez, P, Goy, C, Jezequel, S, Lees, J, Lucotte, A, Martin, F, Merle, E, Minard, M, Nief, J, Odier, P, Pietrzyk, B, Trocme, B, Bravo, S, Casado, M, Chmeissani, M, Comas, P, Crespo, J, Fernandez, E, Fernandez Bosman, M, Garrido, L, Grauges, E, Juste, A, Martinez, M, Merino, G, Miquel, R, Mir, L, Orteu, S, Pacheco, A, Park, I, Perlas, J, Riu, I, Ruiz, H, Sanchez, F, Colaleo, A, Creanza, D, De Filippis, N, De Palma, M, Iaselli, G, Maggi, G, Maggi, M, Nuzzo, S, Ranieri, A, Raso, G, Ruggieri, F, Selvaggi, G, Silvestris, L, Tempesta, P, Tricomi, A, Zito, G, Huang, X, Lin, J, Ouyang, Q, Wang, T, Xie, Y, Xu, R, Xue, S, Zhang, J, Zhang, L, Zhao, W, Abbaneo, D, Bazarko, A, Becker, U, Boix, G, Bird, F, Blucher, E, Bonvicini, B, Bright Thomas, P, Barklow, T, Buchmuller, O, Cattaneo, M, Cerutti, F, Ciulli, V, Clerbaux, B, Drevermann, H, Forty, R, Frank, M, Greening, T, Hagelberg, R, Halley, A, Gianotti, F, Girone, M, Hansen, J, Harvey, J, Jacobsen, R, Hutchcroft, D, Janot, R, Jost, B, Knobloch, J, Kado, M, Lehraus, I, Lazeyras, P, Maley, R, Mato, P, May, J, Moutussi, A, Pepe Altarelli, M, Ranjard, F, Rolandi, L, Schlatter, D, Schmitt, B, Schneider, O, Tejessy, W, Teubert, F, Tomalin, I, Tournefier, E, Veenhof, R, Valassi, A, Wiedenmann, W, Wright, A, Ajaltouni, Z, Badaud, F, Chazelle, G, Deschamps, O, Dessagne, S, Falvard, A, Ferdi, C, Fayolle, D, Gay, P, Guicheney, C, Henrard, P, Jousset, J, Michel, B, Monteil, S, Montret, J, Pallin, D, Pascolo, J, Perret, P, Podlyski, F, Bertelsen, H, Fernley, T, Hansen, P, Kraan, A, Lindahl, A, Mollerud, R, Nilsson, B, Rensch, B, Waananen, A, Daskalakis, G, Kyriakis, A, Markou, C, Simopoulou, E, Siotis, I, Vayaki, A, Zachariadou, K, Blondel, A, Bonneaud, G, Brient, J, Machefert, E, Rouge, A, Rumpf, M, Swynghedauw, M, Tanaka, R, Verderi, M, Videau, H, Focardi, E, Parrini, G, Corden, M, Georgiopoulos, C, Antonelli, A, Antonelli, M, Bencivenni, G, Bologna, G, Bossi, F, Campana, P, Capon, G, Chiarella, V, Felici, G, Laurelli, P, Mannocchi, G, Murtas, G, Passalacqua, L, Picchi, P, Colrain, P, Ten Have, I, Hughes, I, Kennedy, J, Knowles, I, Lynch, J, Morton, W, Negus, P, O'Shea, V, Raine, C, Reeves, P, Scarr, J, Smith, K, Thompson, A, Turnbull, R, Wasserbaech, S, Cavanaugh, R, Dhamotharan, S, Geweniger, C, Hanke, P, Hansper, G, Hepp, V, Kluge, E, Putzer, A, Sommer, J, Stenzel, H, Tittel, K, Werner, W, Wunsch, M, Beuselinck, R, Binnie, D, Cameron, W, Davies, G, Dornan, P, Goodsir, S, Marinelli, N, Martin, E, Nash, J, Nowell, J, Rutherford, S, Sedgbeer, J, Thompson, J, White, R, Williams, M, Ghete, V, Girtler, P, Kneringer, E, Kuhn, D, Rudolph, G, Bouhova Thacker, E, Bowdery, C, Buck, P, Clarke, D, Ellis, G, Finch, A, Foster, F, Hughes, G, Jones, R, Keemer, N, Pearson, M, Robertson, N, Sloan, T, Smizanska, M, Snow, S, Van Der Aa, O, Delaere, C, Leibenguth, G, Lemaitre, V, Bauerdick, L, Blumenschein, U, Van Gemmeren, P, Giehl, I, Holldorfer, F, Jakobs, K, Kasemann, M, Kayser, F, Kleinknecht, K, Muller, A, Quast, G, Renk, B, Rohne, E, Sander, H, Schmeling, S, Wachsmuth, H, Wanke, R, Zeitnitz, C, Ziegler, T, Aubert, J, Benchouk, C, Bonissent, A, Carr, J, 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Vandalen, Gj, Virtue, Cj, Vokurka, Eh, Vollmer, Cf, Wagner, Dl, Walker, Jp, Ward, Cp, Ward, Dr, Watkins, Pm, Watson, At, Watson, Nk, Whalley, Ma, Wilson, Ja, Wilson, Gw, Winterer, Vh, Wood, Nc, Wyatt, Tr, Zorn, Gt, Allen, Nj, Ash, Ww, Baird, Kg, Band, Hr, Barakat, Mb, Baranko, Gj, Barklow, Tl, Bashindzhagian, Gl, Bauer, Jm, Bazarko, Ao, Benvenuti, Ac, Bilei, Gm, Bower, Gr, Brau, Je, Bugg, Wm, Burnett, Th, Burrows, Pn, Caldwell, Do, Chadwick, Gb, Cohn, Ho, Coller, Ja, Convery, Mr, Cowan, Rf, Coyle, Pa, Coyne, Dg, Damerell, Cj, de Groot, N, de Sangro, R, Dervan, Pj, Dong, Dn, Du, Pyc, Eisenstein, Bi, Fernandez, Jp, Fero, Mj, Friedman, Ji, Garwin, El, Hallewell, Gd, Hart, El, Hitlin, Dg, Huffer, Me, Hughes, Ew, Izen, Jm, Jackson, Dj, Jaros, Ja, Jiang, Zy, Johnson, Jr, Johnson, Ra, Kamyshkov, Ya, Kang, Hj, Kelsey, Mh, Kendall, Hw, Kim, Yd, Krishna, Nm, Labs, Jf, Lauber, Ja, Leith, Dwg, Liu, Mx, Lynch, Hl, Markiewicz, Tw, Mcgowan, Jf, Mckemey, Ak, Meadows, Bt, Mockett, Pm, Moffeit, Kc, 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MCGOWAN, A.K. MCKEMEY, B.T. MEADOWS, R. MESSNER, P.M. MOCKETT, K.C. MOFFEIT, T.B. MOORE, M. MORII, B. MOURS, D. MULLER, G. MUELLER, V. MURZIN, T. NAGAMINE, S. NARITA, U. NAUENBERG, H. NEAL, G. NESOM, M. NUSSBAUM, Y. OHNISHI, N. OISHI, D. ONOPRIENKO, L.S. OSBORNE, R.S. PANVINI, C.H.PARK, H. PARK, T.J. PAVEL, I. PERUZZI, L. PESCARA, M. PICCOLO, L. PIEMONTESE, E. PIERONI, K.T. PITTS, R.J. PLANO, R. PREPOST, C.Y. PRESCOTT, G. PUNKAR, J. QUIGLEY, B.N. RATCLIFF, K. REEVES, T.W. REEVES, J. REIDY, P.L. REINERTSEN, P.E. RENSING, L.S. ROCHESTER, J.E. ROTHBERG, P.C. ROWSON, J.J. RUSSELL, O.H. SAXTON, T. SCHALK, R.H. SCHINDLER, U. SCHNEEKLOTH, B.A. SCHUMM, J. SCHWIENING, A. SEIDEN, S. SEN, V.V. SERBO, L. SERVOLI, M.H. SHAEVITZ, J.T. SHANK, G. SHAPIRO, D.J. SHERDEN, K.D. SHMAKOV, C. SIMOPOULOS, N.B. SINEV, S.R. SMITH, M.B. SMY, J.A. SNYDER, M.D. SOKOLOFF, H. STAENGLE, A. STAHL, P. STAMER, H. STEINER, R. STEINER, M.G. STRAUSS, D. SU, F. SUEKANE, A. SUGIYAMA, A. SUZUKI, S. SUZUKI, M. SWARTZ, A. SZUMILO, T. TAKAHASHI, F.E. TAYLOR, J.J. THALER, J. THOM, E. TORRENCE, A.I. TRANDAFIR, J.D. TURK, T. USHER, J. VA VRA, C. VANNINI, E. VELLA, J.P. VENUTI, R. VERDIER, P.G. VERDINI, D.L. WAGNER, S.R. WAGNER, A.P. WAITE, S. WALSTON, J. WANG, S.J. WATTS, A.W. WEIDEMANN, E.R. WEISS, J.S. WHITAKER, S.L. WHITE, F.J. WICKENS, D.A. WILLIAMS, D.C. WILLIAMS, S.H. WILLIAMS, S. WILLOCQ, R.J. WILSON, W.J. WISNIEWSKI, J.L. WITTLIN, M. WOODS, G.B. WORD, T.R. WRIGHT, J. WYSS, R.K. YAMAMOTO, J.M. YAMARTINO, X.Q. YANG, J. YASHIMA, S.J. YELLIN, C.C.YOUNG, H.YUTA, G. ZAPALAC, R.W. ZDARKO, C. ZEITLIN, J. ZHOU, Centre National de la Recherche Scientifique (CNRS)-École polytechnique (X)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3), Centre National de la Recherche Scientifique (CNRS)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université Paris-Sud - Paris 11 (UP11), Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université Pierre et Marie Curie - Paris 6 (UPMC), Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut Polytechnique de Grenoble - Grenoble Institute of Technology-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3), Laboratoire d'Annecy de Physique des Particules (LAPP/Laboratoire d'Annecy-le-Vieux de Physique des Particules), Centre National de la Recherche Scientifique (CNRS)-Université Savoie Mont Blanc (USMB [Université de Savoie] [Université de Chambéry])-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3), Centre National de la Recherche Scientifique (CNRS)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Aix Marseille Université (AMU), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Institut Polytechnique de Grenoble - Grenoble Institute of Technology-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Collège de France (CdF)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), GRUNEVALD M, NEWALD M, SCHAEL S, BARATE R, BRUNELIE, RE R, BUSKULIC D, DE BONIS I, DECAMP D, GHEZ P, GOY C, JE, ZE, QUEL S, LEES J, LUCOTTE A, MARTIN M, MERLE E, MINARD M, NIEF J, ODIER P, PIETRZYK B, TROCME, BRAVO S, CASADO MP, CHMEISSANI M, COMAS P, CRESPO JM, FERNANDEZ E, FERNANDEZ-BOSMAN M, GARRIDO L, GRAUGES E, JUSTE A, MARTINEZ M, MERINO G, MIQUEL R, MIR LM, ORTEU S, PACHECO A, PARK IC, PERLAS J, RIU I, RUIZ H, SANCHEZ F, COLALEO A, CREANZA D, DE FILIPPIS N, DE PALMA M, IASELLI G, MAGGI G, MAGGI M, NUZZO S, RANIERI, A, RASO, G, RUGGIERI F, SELVAGGI, G, SILVESTRIS L, TEMPESTA P, TRICOMI A, ZITO G, HUANG X, LIN J, OUYANG Q, WANG T, XIE Y, XU R, XUE S, ZHANG J, ZHANG L, ZHAO W, ABBANEO D, BAZARKO A, BECKER U, BOIX G, BIRD F, BLUCHER E, BONVICINI B, BRIGHT-THOMAS P, BARKLOW T, BUCHMU, LLER O, CATTANEO M, CERUTTI F, CIULLI V, CLERBAUX B, DREVERMANN H, FORTY RW, FRANK M, GREENING TC, HAGELBERG R, HALLEY AW, GIANOTTI F, GIRONE M, HANSEN JB, HARVEY J, JACOBSEN J, HUTCHCROFT DE, JANOT P, JOST B, KNOBLOCH J, KADO M, LEHRAUS I, LAZEYRAS P, and MALEY P
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Top quark ,FORWARD-BACKWARD ASYMMETRY ,PARTICLE PHYSICS ,LARGE ELECTRON POSITRON COLLIDER ,ALEPH ,DELPHI ,L3 ,OPAL ,General Physics and Astronomy ,01 natural sciences ,7. Clean energy ,High Energy Physics - Experiment ,Settore FIS/04 - Fisica Nucleare e Subnucleare ,High Energy Physics - Experiment (hep-ex) ,High Energy Physics - Phenomenology (hep-ph) ,electron-positron physics ,[PHYS.HEXP]Physics [physics]/High Energy Physics - Experiment [hep-ex] ,Electroweak interaction ,Physics ,Quantum chromodynamics ,Electron–positron physics ,Electroweak interactions ,Decays of heavy intermediate gauge bosons ,Fermion–antifermion production ,Precision measurements at the Z resonance ,Tests of the Standard Model ,Radiative corrections ,Effective coupling constants ,Neutral weak current ,Z boson ,W boson ,Higgs boson ,Particle physics - Experiment ,Settore FIS/01 - Fisica Sperimentale ,FERMION-PAIR PRODUCTION ,HADRONIC-Z-DECAYS ,TOP-QUARK MASS ,ANGLE BHABHA SCATTERING ,W-BOSON MASS ,CROSS-SECTION ASYMMETRY ,Z-LINE-SHAPE ,SEMILEPTONIC BRANCHING RATIOS ,CARLO EVENT GENERATOR ,decays of heavy intermediate gauge bosons ,effective coupling constants ,electroweak interactions ,fermion-antifermion production ,higgs boson ,neutral weak current ,precision measurements at the z resonance ,radiative corrections ,tests of the standard model ,top quark ,w boson ,z boson ,Radiative correction ,High Energy Physics - Phenomenology ,FIS/01 - FISICA SPERIMENTALE ,Física nuclear ,Neutrino ,Particle physics ,FOS: Physical sciences ,ddc:500.2 ,Elementary particle physics ,LEP ,electroweak ,Decays of heavy intermediate gauge boson ,Effective coupling constant ,Partícules (Física nuclear) ,Standard Model ,electroweak theory, Z boson, DELPHI, ALEPH, OPAL, L3 ,0103 physical sciences ,010306 general physics ,Coupling constant ,010308 nuclear & particles physics ,High Energy Physics::Phenomenology ,Fermion ,FORWARD-BACKWARD ASYMMETRY, FERMION-PAIR PRODUCTION, HADRONIC-Z-DECAYS, TOP-QUARK MASS, ANGLE BHABHA SCATTERING, W-BOSON MASS, CROSS-SECTION ASYMMETRY, Z-LINE-SHAPE, SEMILEPTONIC BRANCHING RATIOS, CARLO EVENT GENERATOR ,[PHYS.HPHE]Physics [physics]/High Energy Physics - Phenomenology [hep-ph] ,Experimental High Energy Physics ,Electron–positron physic ,High Energy Physics::Experiment ,FIS/04 - FISICA NUCLEARE E SUBNUCLEARE - Abstract
We report on the final electroweak measurements performed with data taken at the Z resonance by the experiments operating at the electron-positron colliders SLC and LEP. The data consist of 17 million Z decays accumulated by the ALEPH, DELPHI, L3 and OPAL experiments at LEP, and 600 thousand Z decays by the SLD experiment using a polarised beam at SLC. The measurements include cross-sections, forward-backward asymmetries and polarised asymmetries. The mass and width of the Z boson, $\MZ$ and $\GZ$, and its couplings to fermions, for example the $\rho$ parameter and the effective electroweak mixing angle, are precisely measured. The number of light neutrino species is determined to be 2.9840+/-0.0082. The results are compared to the predictions of the Standard Model. Electroweak radiative corrections beyond the running of the QED and QCD coupling constants are observed with a significance of five standard deviations, and in agreement with the Standard Model. Of the many Z-pole measurements, the forward-backward asymmetry in b-quark production shows the largest difference with respect to its Standard Model expectation, at the level of 2.8 standard deviations. Through radiative corrections evaluated in the framework of the Standard Model, the masses of the top quark and the W Boson are predicted. These indirect constraints are compared to the direct measurements, providing a stringent test of the Standard Model. Using in addition the direct measurements of $\Mt$ and $\MW$, the mass of the as yet unobserved Standard Model Higgs boson is predicted., Comment: 302 pages, v2: minor corrections and updates of references. Accepted for publication by Physics Reports, v3: further small corrections and journal version
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- 2006
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4. Low-level radiocaesium exposure alters gene expression in roots of Arabidopsis
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Sahr, T., Voigt, G., Schimmack, W., Paretzke, H.G., and Ernst, D.
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abiotic stress ,Arabidopsis thaliana ,cDNA ,gene expression ,radiation ,radiocaesium ,suppression substractive hybridization (SSH) - Published
- 2005
5. Caesium-affected gene expression in Arabidopsis thaliana
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Sahr, T., Voigt, G., Paretzke, H.G., Schramel, P., and Ernst, D.
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abiotic stress ,Arabidopsis thaliana ,caesium(Cs) ,cDNA ,gene expression ,suppression-subtractive hybridization (SSH) - Published
- 2005
6. Untersuchungen zur Transkriptionsänderung in Arabidopsis thaliana nach Cäsium- und Radiocäsium-Applikation
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Sahr, T.
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Caesium ,Radiocaesium ,Stress ,Array ,Subtraktive Suppressions Hybridisierung ,Caesium-Aufnahme ,Subtraktive Suppression Hybridisation ,Caesium uptake - Abstract
Anhand des Transkriptionsmusters von zahlreichen Stressgenen wurde die Wirkungsweise von Cäsium und ionisierender Strahlung auf den Stoffwechsel von Arabidopsis thaliana charakterisiert Die Kombination der Subtraktiven-Suppressions-Hybridisierung und der Makroarray-Technologie erwies sich als eine sehr sensitive Methode, um eine Vielzahl von differentiell exprimierten Genen mit hoher Reproduzierbarkeit und hohem Durchsatz zu analysieren; mit der quantitativen und semiquantitativen PCR konnten schwach induzierte oder reprimierte Gene effektiv verifiziert werden. Cäsium- und Kalium-Ionen besitzen große chemische Gemeinsamkeiten. Während Kalium jedoch für die Pflanze essentiell ist, besitzt Cäsium keine bekannten physiologischen Funktionen. Folgende Reaktionen der Pflanze auf erhöhte Cäsium-133 Konzentrationen (150µM) in der Umgebung konnten auf Transkriptebene nachgewiesen werden: Eine verstärkte Transkription von Redoxenzymen (Superoxid-Dismutase, Thioredoxin, Peroxidase) und die Induktion der Schwefelaufnahme bzw. die Synthese schwefelhaltiger Verbindungen. Eine Induktion und Repression von Enzymen auf Genebene, die sowohl bei der Stabilisierung (Chaperone, Heat-shock Proteine) als auch beim Abbau von Proteinen (Peptidasen, Protease-Inhibitoren) eine Rolle spielen. Eine erhöhte Transkriptionsrate von Genen, deren Produkte als Schutzstoffe für Makromoleküle oder als osmotisch wirksame Substanzen fungieren können (Prolin, Osmotin, LEA-Proteine, etc.); parallel wurde die Expressionsrate verschiedener Aquaporin-Gene verringert. Radiocäsium (Cäsium-134) wurde nur in sehr geringen Konzentrationen und Aktivitäten (30Bq/cm3) in den Versuchen eingesetzt, da vor allem die Langzeitwirkung schwacher ionisierender Strahlung auf Genebene untersucht werden sollte. Dementsprechend waren deutliche Unterschiede in der Stressantwort der Pflanze im Vergleich zu den Cäsium-133 Experimenten zu erkennen: Verstärkte Transkription von Genen des Excisions-Reparatur-Systems und der Rekombination-Reparatur. Beeinflussung der Zellteilung durch Induktion des Gen des CylinB1 und anderer Zellzyklus-kontrollierender Gene. Zudem wurden einige Enzyme des Cytoskelett-Stoffwechsels auf Transkriptionsebene beeinträchtigt (Profilin2, putativer Actin-depolymerisierender Faktor, beta-5 Tubulin). Analog zu den Cäsium-133 Versuchen wurde eine Induktion von Enzymen des oxidativen Stresses gefunden (Superoxid-Dismutase, Thioredoxin, Katalase). Aufnahmeexperimente mit Cäsium und Radiocäsium zeigten, dass das Alkalimetall in der Pflanze in Abhängigkeit zur externen Konzentration akkumuliert wurde; gleichzeitig nahm die Kaliumkonzentration in Arabidopsis ab. Bei hohen Cäsiumkonzentrationen traten an den Blättern Welkeerscheinungen und nekrotische Veränderungen auf. Zudem wurde bei Arabidopsis thaliana das Wachstum von Wurzeln und später von Blättern reduziert. Diese Symptome waren dabei stark von der Kaliumkonzentration im Medium abhängig, weshalb der toxische Schwellenwert als Cäsium/Kalium-Verhältnis angegeben werden sollte. Schwache Strahlung hatte keine Auswirkungen auf den Phänotypen. Erst bei Aktivitäten von 60Bq/cm3 im Medium wurde ein reduziertes Wachstum festgestellt. Ein Vergleich verschiedener Kalium-Transportermutanten mit dem Wildtyp ergab keine Erkenntnisse über das Aufnahmeverhalten von Cäsium. Ausnahme hiervon war die csi52-Mutante, bei der eine leichte Reduction der Cäsium-Aufnahme beobachtet werden konnte; parallel dazu wurde auf Genebene eine verstärkte Transkription verschiedener Kaliumkanäle (KCO, ATKC) im Vergleich zum Wildtyp festgestellt. Zur Klärung des Phänomens wurden erste Versuche unternommen, Gene verschiedener Kanäle in Arabidopsis thaliana überzuexprimieren bzw. auszuschalten (RNAi).
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- 2003
7. Signal transduction pathway in differently ozone-sensitive poplar plants
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Diara, C., Sebastiani, Luca, Langebartels, C., Sahr, T., Mensuali, Anna, Baldan, B., and Ranieri, A.
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- 2003
8. INDUZIONE DI MOLECOLE SEGNALE IN RISPOSTA AD UN'ESPOSIZIONE ACUTA ALL'O3 IN PIANTE DI PIOPPO
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Diara, Cecilia, Sebastiani, L., Vitagliano, C., Langebartels, C., Sahr, T., Soldatini, Gianfranco, and Ranieri, Annamaria
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- 2003
9. Induzione di molecole segnale in risposta ad un’esposizione acuta dell’O3 in piante di pioppo
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Diara, C., Sebastiani, Luca, Vitagliano, Claudio, Langebartkets, C., Sahr, T., Soldatini, G. F., and Ranieri, A.
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- 2002
10. The Legionella pneumophila F-box protein Lpp2082 (AnkB) modulates ubiquitination of the host protein parvin B and promotes intracellular replication
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Lomma, M., primary, Dervins-Ravault, D., additional, Rolando, M., additional, Nora, T., additional, Newton, H. J., additional, Sansom, F. M., additional, Sahr, T., additional, Gomez-Valero, L., additional, Jules, M., additional, Hartland, E. L., additional, and Buchrieser, C., additional
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- 2010
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11. Tetrahydrofolate biosynthesis and distribution in higher plants
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Sahr, T., primary, Ravanel, S., additional, and Rébeillé, F., additional
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- 2005
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12. Mechanistic and inhibition studies of chorismate-utilizing enzymes
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Kerbarh, O., primary, Bulloch, E.M.M., additional, Payne, R.J., additional, Sahr, T., additional, Rébeillé, F., additional, and Abell, C., additional
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- 2005
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13. The T4bSS of Legionella features a two-step secretion pathway with an inner membrane intermediate for secretion of transmembrane effectors.
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Malmsheimer S, Grin I, Bohn E, Franz-Wachtel M, Macek B, Sahr T, Smollich F, Chetrit D, Meir A, Roy C, Buchrieser C, and Wagner S
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- Cell Membrane metabolism, Secretory Pathway physiology, Membrane Proteins metabolism, Bacterial Proteins metabolism, Legionella pneumophila metabolism, Type IV Secretion Systems metabolism, Protein Transport physiology
- Abstract
To promote intracellular survival and infection, Legionella spp. translocate hundreds of effector proteins into eukaryotic host cells using a type IV b protein secretion system (T4bSS). T4bSS are well known to translocate soluble as well as transmembrane domain-containing effector proteins (TMD-effectors) but the mechanisms of secretion are still poorly understood. Herein we investigated the secretion of hydrophobic TMD-effectors, of which about 80 were previously reported to be encoded by L. pneumophila. A proteomic analysis of fractionated membranes revealed that TMD-effectors are targeted to and inserted into the bacterial inner membranes of L. pneumophila independent of the presence of a functional T4bSS. While the T4bSS chaperones IcmS and IcmW were critical for secretion of all tested TMD-effectors, they did not influence inner membrane targeting of these proteins. As for soluble effector proteins, translocation of all investigated TMD-effectors depended on a C-terminal secretion signal. A deeper analysis of the TMD-effector SidF showed that this signal needed to be presented towards the cytoplasmic side of the inner membrane and that a small periplasmic loop was required for efficient translocation. We propose that strongly hydrophobic TMD-effectors are secreted in a two-step secretion process: Initially, an inner membrane intermediate is formed, that is extracted towards the cytoplasmic side, possibly by the help of the type IV coupling protein complex and subsequently secreted into eukaryotic host cells by the T4bSS core complex. Overall, our study highlights the amazing versatility of T4bSS to secrete soluble and TMD-effectors from different subcellular locations of the bacterial cell., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Malmsheimer et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
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- 2024
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14. Structural basis for the toxicity of Legionella pneumophila effector SidH.
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Sharma R, Adams M, Griffith-Jones S, Sahr T, Gomez-Valero L, Weis F, Hons M, Gharbi S, Berkane R, Stolz A, Buchrieser C, and Bhogaraju S
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- Humans, Escherichia coli metabolism, Peptide Elongation Factor Tu metabolism, Ubiquitin-Protein Ligases metabolism, Bacterial Proteins metabolism, Legionella pneumophila metabolism, Legionnaires' Disease
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Legionella pneumophila (LP) secretes more than 300 effectors into the host cytosol to facilitate intracellular replication. One of these effectors, SidH, 253 kDa in size with no sequence similarity to proteins of known function is toxic when overexpressed in host cells. SidH is regulated by the LP metaeffector LubX which targets SidH for degradation in a temporal manner during LP infection. The mechanism underlying the toxicity of SidH and its role in LP infection are unknown. Here, we determined the cryo-EM structure of SidH at 2.7 Å revealing a unique alpha helical arrangement with no overall similarity to known protein structures. Surprisingly, purified SidH came bound to a E. coli EF-Tu/t-RNA/GTP ternary complex which could be modeled into the cryo-EM density. Mutation of residues disrupting the SidH-tRNA interface and SidH-EF-Tu interface abolish the toxicity of overexpressed SidH in human cells, a phenotype confirmed in infection of Acanthamoeba castellani. We also present the cryo-EM structure of SidH in complex with a U-box domain containing ubiquitin ligase LubX delineating the mechanism of regulation of SidH. Our data provide the basis for the toxicity of SidH and into its regulation by the metaeffector LubX., (© 2023. The Author(s).)
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- 2023
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15. Chromosome-scale assemblies of Acanthamoeba castellanii genomes provide insights into Legionella pneumophila infection-related chromatin reorganization.
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Matthey-Doret C, Colp MJ, Escoll P, Thierry A, Moreau P, Curtis B, Sahr T, Sarrasin M, Gray MW, Lang BF, Archibald JM, Buchrieser C, and Koszul R
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- Chromatin Assembly and Disassembly, Genome, Protozoan, Chromatin metabolism, Chromatin genetics, Legionnaires' Disease microbiology, Humans, Acanthamoeba castellanii microbiology, Acanthamoeba castellanii genetics, Legionella pneumophila genetics, Legionella pneumophila pathogenicity
- Abstract
The unicellular amoeba Acanthamoeba castellanii is ubiquitous in aquatic environments, where it preys on bacteria. The organism also hosts bacterial endosymbionts, some of which are parasitic, including human pathogens such as Chlamydia and Legionella spp. Here we report complete, high-quality genome sequences for two extensively studied A. castellanii strains, Neff and C3. Combining long- and short-read data with Hi-C, we generated near chromosome-level assemblies for both strains with 90% of the genome contained in 29 scaffolds for the Neff strain and 31 for the C3 strain. Comparative genomics revealed strain-specific functional enrichment, most notably genes related to signal transduction in the C3 strain and to viral replication in Neff. Furthermore, we characterized the spatial organization of the A. castellanii genome and showed that it is reorganized during infection by Legionella pneumophila Infection-dependent chromatin loops were found to be enriched in genes for signal transduction and phosphorylation processes. In genomic regions where chromatin organization changed during Legionella infection, we found functional enrichment for genes associated with metabolism, organelle assembly, and cytoskeleton organization. Given Legionella infection is known to alter its host's cell cycle, to exploit the host's organelles, and to modulate the host's metabolism in its favor, these changes in chromatin organization may partly be related to mechanisms of host control during Legionella infection., (© 2022 Matthey-Doret et al.; Published by Cold Spring Harbor Laboratory Press.)
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- 2022
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16. Translocated Legionella pneumophila small RNAs mimic eukaryotic microRNAs targeting the host immune response.
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Sahr T, Escoll P, Rusniok C, Bui S, Pehau-Arnaudet G, Lavieu G, and Buchrieser C
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- Bacterial Proteins metabolism, Cell Line, DEAD Box Protein 58, Eukaryota genetics, Extracellular Vesicles, Humans, Immunity, Innate, Interleukin-1 Receptor-Associated Kinases, Legionnaires' Disease microbiology, Receptors, Immunologic, Signal Transduction, Eukaryota immunology, Host-Pathogen Interactions immunology, Legionella pneumophila metabolism, Legionnaires' Disease immunology, MicroRNAs genetics, MicroRNAs metabolism
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Legionella pneumophila is an intracellular bacterial pathogen that can cause a severe form of pneumonia in humans, a phenotype evolved through interactions with aquatic protozoa in the environment. Here, we show that L. pneumophila uses extracellular vesicles to translocate bacterial small RNAs (sRNAs) into host cells that act on host defence signalling pathways. The bacterial sRNA RsmY binds to the UTR of ddx58 (RIG-I encoding gene) and cRel, while tRNA-Phe binds ddx58 and irak1 collectively reducing expression of RIG-I, IRAK1 and cRel, with subsequent downregulation of IFN-β. Thus, RsmY and tRNA-Phe are bacterial trans-kingdom regulatory RNAs downregulating selected sensor and regulator proteins of the host cell innate immune response. This miRNA-like regulation of the expression of key sensors and regulators of immunity is a feature of L. pneumophila host-pathogen communication and likely represents a general mechanism employed by bacteria that interact with eukaryotic hosts., (© 2022. The Author(s).)
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- 2022
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17. Reverting the mode of action of the mitochondrial F O F 1 -ATPase by Legionella pneumophila preserves its replication niche.
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Escoll P, Platon L, Dramé M, Sahr T, Schmidt S, Rusniok C, and Buchrieser C
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- Adenosine Triphosphate genetics, Bacterial Proteins metabolism, Legionella pneumophila genetics, Mitochondria genetics, Mitochondrial Proteins genetics, Mitochondrial Proteins metabolism, Proton-Translocating ATPases metabolism, Type IV Secretion Systems metabolism, Adenosine Triphosphate metabolism, Bacterial Proteins genetics, Legionella pneumophila metabolism, Mitochondria metabolism, Proton-Translocating ATPases deficiency
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Legionella pneumophila, the causative agent of Legionnaires' disease, a severe pneumonia, injects via a type 4 secretion system (T4SS) more than 300 proteins into macrophages, its main host cell in humans. Certain of these proteins are implicated in reprogramming the metabolism of infected cells by reducing mitochondrial oxidative phosphorylation (OXPHOS) early after infection. Here. we show that despite reduced OXPHOS, the mitochondrial membrane potential (Δ ψ
m ) is maintained during infection of primary human monocyte-derived macrophages (hMDMs). We reveal that L. pneumophila reverses the ATP-synthase activity of the mitochondrial FO F1 -ATPase to ATP-hydrolase activity in a T4SS-dependent manner, which leads to a conservation of the Δ ψm , preserves mitochondrial polarization, and prevents macrophage cell death. Analyses of T4SS effectors known to target mitochondrial functions revealed that Lp Spl is partially involved in conserving the Δ ψm , but not LncP and MitF. The inhibition of the L. pneumophila -induced 'reverse mode' of the FO F1 -ATPase collapsed the Δ ψm and caused cell death in infected cells. Single-cell analyses suggested that bacterial replication occurs preferentially in hMDMs that conserved the Δ ψm and showed delayed cell death. This direct manipulation of the mode of activity of the FO F1 -ATPase is a newly identified feature of L. pneumophila allowing to delay host cell death and thereby to preserve the bacterial replication niche during infection., Competing Interests: PE, LP, MD, TS, SS, CR, CB No competing interests declared, (© 2021, Escoll et al.)- Published
- 2021
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18. [Results of the systematic literature search as basis for the "Evidence-based treatment recommendations for familial Mediterranean fever patients with insufficient response or intolerability to colchicine" of the Society for Pediatric and Adolescent Rheumatology and the German Society for Rheumatology].
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Sahr T, Kiltz U, Weseloh C, Kallinich T, and Braun J
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- Adolescent, Adult, Child, Colchicine adverse effects, Colchicine therapeutic use, Germany, Humans, Biological Products, Familial Mediterranean Fever diagnosis, Familial Mediterranean Fever drug therapy, Rheumatology
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Background: Familial Mediterranean fever (FMF) is a genetic disease of childhood and adulthood which is relatively rare in Germany. It is characterized by recurrent febrile attacks, peritonitis, pleuritis and arthritis. The established treatment with colchicine is effective and well-tolerated by most patients; however, some patients do not adequately respond or do not tolerate this treatment. Biologics can be considered for some of these patients. The Society for Pediatric and Adolescent Rheumatology (GKJR) and the German Society for Rheumatology (DGRh) have agreed to develop joint recommendations for this specific clinical situation., Aim: Implementation of a systematic literature search (SLR) on the basis of the EULAR recommendations published in 2016 as the foundation for the development of evidence-based treatment recommendations for FMF patients with insufficient response or intolerance to colchicine., Methods: The SLR was performed using references from various databases as an update of the SLR carried out by EULAR up to 2014, whereby all articles must have been published between 1 January 2015 and 31 December 2017. The Rayyan abstract tool for the preselection and the classification of the Oxford Centre for Evidence Based Medicine 2009 were used for the preparation of the evidence tables., Results: The search yielded 360 hits and after duplicate matching 263. A total of 88 publications were included (34%) and 102 excluded (39%), a review of the full publication was necessary for a further 73 (28%) and 43 were discussed more intensively. Finally, 64 publications (24%) remained. A total of 4 case-control studies, 31 cohort studies, 8 case series, 7 controlled studies (including 5 abstracts), 10 reviews, 4 meta-analyses and systematic reviews were accepted., Discussion: The SLR was carried out in a scientifically accurate and transparent manner according to international standards. The SLR proved to be a good basis for a consensus on the 5 overarching principles and the 10 recommendations, so that the joint activity of the GKJR and DGRh was successfully and even promptly concluded. The recommendations are a solid basis for treating patients of all ages with FMF. The explanations on the problem of colchicine resistance play an important role here.
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- 2020
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19. Improving representation in telephone-based health care access surveys requires purposeful efforts to include prepaid cell phone users.
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Berzofsky ME, Scruggs CB, Lu B, Speizer H, and Sahr T
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- Cell Phone economics, Humans, Patient Protection and Affordable Care Act, Socioeconomic Factors, Surveys and Questionnaires, United States, Cell Phone statistics & numerical data, Health Services Accessibility statistics & numerical data, Telemedicine statistics & numerical data
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- 2019
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20. The pleiotropic Legionella transcription factor LvbR links the Lqs and c-di-GMP regulatory networks to control biofilm architecture and virulence.
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Hochstrasser R, Kessler A, Sahr T, Simon S, Schell U, Gomez-Valero L, Buchrieser C, and Hilbi H
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- 4-Butyrolactone analogs & derivatives, Bacterial Proteins genetics, Cyclic GMP metabolism, Legionella pneumophila pathogenicity, Legionnaires' Disease microbiology, Quorum Sensing, Virulence, Bacterial Proteins metabolism, Biofilms, Cyclic GMP analogs & derivatives, Gene Expression Regulation, Bacterial, Gene Regulatory Networks, Legionella pneumophila genetics, Transcription Factors metabolism
- Abstract
The causative agent of Legionnaires' disease, Legionella pneumophila, colonizes amoebae and biofilms in the environment. The opportunistic pathogen employs the Lqs (Legionella quorum sensing) system and the signalling molecule LAI-1 (Legionella autoinducer-1) to regulate virulence, motility, natural competence and expression of a 133 kb genomic "fitness island", including a putative novel regulator. Here, we show that the regulator termed LvbR is an LqsS-regulated transcription factor that binds to the promoter of lpg1056/hnox1 (encoding an inhibitor of the diguanylate cyclase Lpg1057), and thus, regulates proteins involved in c-di-GMP metabolism. LvbR determines biofilm architecture, since L. pneumophila lacking lvbR accumulates less sessile biomass and forms homogeneous mat-like structures, while the parental strain develops more compact bacterial aggregates. Comparative transcriptomics of sessile and planktonic ΔlvbR or ΔlqsR mutant strains revealed concerted (virulence, fitness island, metabolism) and reciprocally (motility) regulated genes in biofilm and broth respectively. Moreover, ΔlvbR is hyper-competent for DNA uptake, defective for phagocyte infection, outcompeted by the parental strain in amoebae co-infections and impaired for cell migration inhibition. Taken together, our results indicate that L. pneumophila LvbR is a novel pleiotropic transcription factor, which links the Lqs and c-di-GMP regulatory networks to control biofilm architecture and pathogen-host cell interactions., (© 2019 Society for Applied Microbiology and John Wiley & Sons Ltd.)
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- 2019
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21. The Life Cycle of L. pneumophila : Cellular Differentiation Is Linked to Virulence and Metabolism.
- Author
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Oliva G, Sahr T, and Buchrieser C
- Subjects
- Animals, Energy Metabolism, Environment, Humans, Legionnaires' Disease immunology, Legionnaires' Disease metabolism, Metabolic Networks and Pathways, Virulence, Host-Pathogen Interactions immunology, Legionella pneumophila physiology, Legionnaires' Disease microbiology, Life Cycle Stages
- Abstract
Legionella pneumophila is a gram-negative bacterium that inhabits freshwater ecosystems, where it is present in biofilm or as planktonic form. L. pneumophila is mainly found associated with protozoa, which serve as protection from hostile environments and as replication niche. If inhaled within aerosols, L. pneumophila is also able to infect and replicate in human alveolar macrophages, eventually causing the Legionnaires' disease. The transition between intracellular and extracellular environments triggers a differentiation program in which metabolic as well as morphogenetic changes occur. We here describe the current knowledge on how the different developmental states of this bacterium are regulated, with a particular emphasis on the stringent response activated during the transition from the replicative phase to the infectious phase and the metabolic features going in hand. We propose that the cellular differentiation of this intracellular pathogen is closely associated to key metabolic changes in the bacterium and the host cell, which together have a crucial role in the regulation of L. pneumophila virulence.
- Published
- 2018
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22. Legionella pneumophila CsrA regulates a metabolic switch from amino acid to glycerolipid metabolism.
- Author
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Häuslein I, Sahr T, Escoll P, Klausner N, Eisenreich W, and Buchrieser C
- Subjects
- Cell Respiration, Citric Acid Cycle, Glucose metabolism, Glycolysis, Legionella pneumophila metabolism, Pentose Phosphate Pathway, Amino Acids metabolism, Bacterial Proteins metabolism, Glycolipids metabolism, Repressor Proteins metabolism
- Abstract
Legionella pneumophila CsrA plays a crucial role in the life-stage-specific expression of virulence phenotypes and metabolic activity. However, its exact role is only partly known. To elucidate how CsrA impacts L. pneumophila metabolism we analysed the CsrA depended regulation of metabolic functions by comparative
13 C-isotopologue profiling and oxygen consumption experiments of a L. pneumophila wild-type (wt) strain and its isogenic csrA- mutant. We show that a csrA- mutant has significantly lower respiration rates when serine, alanine, pyruvate, α-ketoglutarate or palmitate is the sole carbon source. By contrast, when grown in glucose or glycerol, no differences in respiration were detected. Isotopologue profiling uncovered that the transfer of label from [U-13 C3 ]serine via pyruvate into the citrate cycle and gluconeogenesis was lower in the mutant as judged from the labelling patterns of protein-derived amino acids, cell-wall-derived diaminopimelate, sugars and amino sugars and 3-hydroxybutyrate derived from polyhydroxybutyrate (PHB). Similarly, the incorporation of [U-13 C6 ]glucose via the glycolysis/Entner-Doudoroff (ED) pathway but not via the pentose phosphate pathway was repressed in the csrA- mutant. On the other hand, fluxes due to [U-13 C3 ]glycerol utilization were increased in the csrA- mutant. In addition, we showed that exogenous [1,2,3,4-13 C4 ]palmitic acid is efficiently used for PHB synthesis via13 C2 -acetyl-CoA. Taken together, CsrA induces serine catabolism via the tricarboxylic acid cycle and glucose degradation via the ED pathway, but represses glycerol metabolism, fatty acid degradation and PHB biosynthesis, in particular during exponential growth. Thus, CsrA has a determining role in substrate usage and carbon partitioning during the L. pneumophila life cycle and regulates a switch from amino acid usage in replicative phase to glycerolipid usage during transmissive growth., (© 2017 The Authors.)- Published
- 2017
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23. The Legionella pneumophila genome evolved to accommodate multiple regulatory mechanisms controlled by the CsrA-system.
- Author
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Sahr T, Rusniok C, Impens F, Oliva G, Sismeiro O, Coppée JY, and Buchrieser C
- Subjects
- Bacterial Proteins metabolism, Base Sequence, Blotting, Northern, Evolution, Molecular, Feedback, Physiological, Gene Expression Profiling methods, Glycolysis genetics, Host-Pathogen Interactions, Humans, Legionella pneumophila metabolism, Legionella pneumophila pathogenicity, Legionnaires' Disease microbiology, Mutation, Oligonucleotide Array Sequence Analysis, Operon genetics, Pentose Phosphate Pathway genetics, Protein Binding, RNA, Messenger genetics, RNA, Messenger metabolism, Repressor Proteins metabolism, Riboswitch genetics, Tandem Mass Spectrometry, Virulence genetics, Bacterial Proteins genetics, Gene Expression Regulation, Bacterial, Genome, Bacterial genetics, Legionella pneumophila genetics, Repressor Proteins genetics
- Abstract
The carbon storage regulator protein CsrA regulates cellular processes post-transcriptionally by binding to target-RNAs altering translation efficiency and/or their stability. Here we identified and analyzed the direct targets of CsrA in the human pathogen Legionella pneumophila. Genome wide transcriptome, proteome and RNA co-immunoprecipitation followed by deep sequencing of a wild type and a csrA mutant strain identified 479 RNAs with potential CsrA interaction sites located in the untranslated and/or coding regions of mRNAs or of known non-coding sRNAs. Further analyses revealed that CsrA exhibits a dual regulatory role in virulence as it affects the expression of the regulators FleQ, LqsR, LetE and RpoS but it also directly regulates the timely expression of over 40 Dot/Icm substrates. CsrA controls its own expression and the stringent response through a regulatory feedback loop as evidenced by its binding to RelA-mRNA and links it to quorum sensing and motility. CsrA is a central player in the carbon, amino acid, fatty acid metabolism and energy transfer and directly affects the biosynthesis of cofactors, vitamins and secondary metabolites. We describe the first L. pneumophila riboswitch, a thiamine pyrophosphate riboswitch whose regulatory impact is fine-tuned by CsrA, and identified a unique regulatory mode of CsrA, the active stabilization of RNA anti-terminator conformations inside a coding sequence preventing Rho-dependent termination of the gap operon through transcriptional polarity effects. This allows L. pneumophila to regulate the pentose phosphate pathway and the glycolysis combined or individually although they share genes in a single operon. Thus the L. pneumophila genome has evolved to acclimate at least five different modes of regulation by CsrA giving it a truly unique position in its life cycle.
- Published
- 2017
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24. A Unique cis-Encoded Small Noncoding RNA Is Regulating Legionella pneumophila Hfq Expression in a Life Cycle-Dependent Manner.
- Author
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Oliva G, Sahr T, Rolando M, Knoth M, and Buchrieser C
- Subjects
- Acanthamoeba castellanii microbiology, Gene Deletion, Host Factor 1 Protein genetics, Legionella pneumophila genetics, RNA, Small Untranslated genetics, Virulence, Gene Expression Regulation, Bacterial, Host Factor 1 Protein biosynthesis, Legionella pneumophila growth & development, Legionella pneumophila metabolism, RNA, Small Untranslated metabolism
- Abstract
Legionella pneumophila is an environmental bacterium that parasitizes protozoa, but it may also infect humans, thereby causing a severe pneumonia called Legionnaires' disease. To cycle between the environment and a eukaryotic host, L. pneumophila is regulating the expression of virulence factors in a life cycle-dependent manner: replicating bacteria do not express virulence factors, whereas transmissive bacteria are highly motile and infective. Here we show that Hfq is an important regulator in this network. Hfq is highly expressed in transmissive bacteria but is expressed at very low levels in replicating bacteria. A L. pneumophila hfq deletion mutant exhibits reduced abilities to infect and multiply in Acanthamoeba castellanii at environmental temperatures. The life cycle-dependent regulation of Hfq expression depends on a unique cis-encoded small RNA named Anti-hfq that is transcribed antisense of the hfq transcript and overlaps its 5' untranslated region. The Anti-hfq sRNA is highly expressed only in replicating L. pneumophila where it regulates hfq expression through binding to the complementary regions of the hfq transcripts. This results in reduced Hfq protein levels in exponentially growing cells. Both the small noncoding RNA (sRNA) and hfq mRNA are bound and stabilized by the Hfq protein, likely leading to the cleavage of the RNA duplex by the endoribonuclease RNase III. In contrast, after the switch to transmissive bacteria, the sRNA is not expressed, allowing now an efficient expression of the hfq gene and consequently Hfq. Our results place Hfq and its newly identified sRNA anti-hfq in the center of the regulatory network governing L. pneumophila differentiation from nonvirulent to virulent bacteria., Importance: The abilities of L. pneumophila to replicate intracellularly and to cause disease depend on its capacity to adapt to different extra- and intracellular environmental conditions. Therefore, a timely and fine-tuned expression of virulence factors and adaptation traits is crucial. Yet, the regulatory circuits governing the life cycle of L. pneumophila from replicating to virulent bacteria are only partly uncovered. Here we show that the life cycle-dependent regulation of the RNA chaperone Hfq relies on a small regulatory RNA encoded antisense to the hfq-encoding gene through a base pairing mechanism. Furthermore, Hfq regulates its own expression in an autoregulatory loop. The discovery of this RNA regulatory mechanism in L. pneumophila is an important step forward in the understanding of how the switch from inoffensive, replicating to highly virulent, transmissive L. pneumophila is regulated., (Copyright © 2017 Oliva et al.)
- Published
- 2017
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25. The α-hydroxyketone LAI-1 regulates motility, Lqs-dependent phosphorylation signalling and gene expression of Legionella pneumophila.
- Author
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Schell U, Simon S, Sahr T, Hager D, Albers MF, Kessler A, Fahrnbauer F, Trauner D, Hedberg C, Buchrieser C, and Hilbi H
- Subjects
- Alkanes pharmacology, Cell Movement, Escherichia coli genetics, Gene Expression, Gene Expression Profiling, Gene Expression Regulation, Bacterial, Host-Pathogen Interactions, Ketones pharmacology, Legionella pneumophila drug effects, Legionella pneumophila growth & development, Movement, Phosphorylation, Quorum Sensing, Transcription Factors metabolism, Vibrio cholerae genetics, Alkanes metabolism, Ketones metabolism, Legionella pneumophila genetics, Legionella pneumophila metabolism, Signal Transduction genetics
- Abstract
The causative agent of Legionnaires' disease, Legionella pneumophila, employs the autoinducer compound LAI-1 (3-hydroxypentadecane-4-one) for cell-cell communication. LAI-1 is produced and detected by the Lqs (Legionella quorum sensing) system, comprising the autoinducer synthase LqsA, the sensor kinases LqsS and LqsT, as well as the response regulator LqsR. Lqs-regulated processes include pathogen-host interactions, production of extracellular filaments and natural competence for DNA uptake. Here we show that synthetic LAI-1 promotes the motility of L. pneumophila by signalling through LqsS/LqsT and LqsR. Upon addition of LAI-1, autophosphorylation of LqsS/LqsT by [γ-(32) P]-ATP was inhibited in a dose-dependent manner. In contrast, the Vibrio cholerae autoinducer CAI-1 (3-hydroxytridecane-4-one) promoted the phosphorylation of LqsS (but not LqsT). LAI-1 did neither affect the stability of phospho-LqsS or phospho-LqsT, nor the dephosphorylation by LqsR. Transcriptome analysis of L. pneumophila treated with LAI-1 revealed that the compound positively regulates a number of genes, including the non-coding RNAs rsmY and rsmZ, and negatively regulates the RNA-binding global regulator crsA. Accordingly, LAI-1 controls the switch from the replicative to the transmissive growth phase of L. pneumophila. In summary, the findings indicate that LAI-1 regulates motility and the biphasic life style of L. pneumophila through LqsS- and LqsT-dependent phosphorylation signalling., (© 2015 John Wiley & Sons Ltd.)
- Published
- 2016
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26. Small RNAs, 5' UTR elements and RNA-binding proteins in intracellular bacteria: impact on metabolism and virulence.
- Author
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Oliva G, Sahr T, and Buchrieser C
- Subjects
- Bacteria genetics, Bacteria metabolism, Bacteria pathogenicity, Gene Expression Regulation, Bacterial, Protein Binding, 5' Untranslated Regions genetics, Bacterial Physiological Phenomena genetics, RNA, Bacterial metabolism, Virulence genetics
- Abstract
Sequencing-based studies have illuminated increased transcriptional complexity within the genome structure of bacteria and have resulted in the identification of many small regulatory RNAs (sRNA) and a large amount of antisense transcription. It remains an open question whether these sRNAs all indeed play regulatory roles, but their identification led to an exponential increase in studies searching for their function. This allowed to show that sRNAs may modulate virulence gene expression, cellular differentiation, metabolic functions, adaptation to environmental conditions and pathogenesis. In this review we will provide mechanistic insights into how sRNAs bind mRNAs and/or proteins. Furthermore, the important roles of the RNA chaperone Hfq, the CsrA system and the CRISPR RNA will be discussed. We will then focus on sRNAs and 5(') untranslated region (UTR) elements of intracellular bacteria like Chlamydia, Listeria, Legionella, or Salmonella, and place emphasis on those that are expressed during replication in host cells and are implicated in virulence and metabolism. In addition, sRNAs that regulate motility, iron homeostasis, and differentiation or stress responses will be highlighted. Taken together sRNAs constitute key elements in many major regulatory networks governing the intracellular life and virulence of pathogenic bacteria., (© FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2015
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27. IroT/mavN, a new iron-regulated gene involved in Legionella pneumophila virulence against amoebae and macrophages.
- Author
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Portier E, Zheng H, Sahr T, Burnside DM, Mallama C, Buchrieser C, Cianciotto NP, and Héchard Y
- Subjects
- Bacterial Proteins metabolism, Base Sequence, Biological Transport, Humans, Legionella pneumophila genetics, Membrane Proteins genetics, Molecular Sequence Data, Virulence, Virulence Factors genetics, Amoeba microbiology, Iron metabolism, Legionella pneumophila metabolism, Legionella pneumophila pathogenicity, Legionnaires' Disease microbiology, Macrophages microbiology, Membrane Proteins metabolism, Virulence Factors metabolism
- Abstract
Legionella pneumophila is a pathogenic bacterium commonly found in water. Eventually, it could be transmitted to humans via inhalation of contaminated aerosols. Iron is known as a key requirement for the growth of L. pneumophila in the environment and within its hosts. Many studies were performed to understand iron utilization by L. pneumophila but no global approaches were conducted. In this study, transcriptomic analyses were performed, comparing gene expression in L. pneumophila in standard versus iron restricted conditions. Among the regulated genes, a newly described one, lpp_2867, was highly induced in iron-restricted conditions. Mutants lacking this gene in L. pneumophila were not affected in siderophore synthesis or utilization. On the contrary, they were defective for growth on iron-depleted solid media and for ferrous iron uptake. A sequence analysis predicts that Lpp_2867 is a membrane protein, suggesting that it is involved in ferrous iron transport. We thus named it IroT, for iron transporter. Infection assays showed that the mutants are highly impaired in intracellular growth within their environmental host Acanthamoeba castellanii and human macrophages. Taken together, our results show that IroT is involved, directly or indirectly, in ferrous iron transport and is a key virulence factor for L. pneumophila., (© 2014 Society for Applied Microbiology and John Wiley & Sons Ltd.)
- Published
- 2015
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28. The Legionella pneumophila kai operon is implicated in stress response and confers fitness in competitive environments.
- Author
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Loza-Correa M, Sahr T, Rolando M, Daniels C, Petit P, Skarina T, Gomez Valero L, Dervins-Ravault D, Honoré N, Savchenko A, and Buchrieser C
- Subjects
- Acanthamoeba castellanii microbiology, Acanthamoeba castellanii physiology, Adaptation, Physiological, Bacterial Proteins genetics, Circadian Clocks, Circadian Rhythm Signaling Peptides and Proteins genetics, Gene Expression Profiling, Gene Expression Regulation, Bacterial, Genetic Fitness, Legionella pneumophila physiology, Phosphorylation, Phylogeny, Protein Structure, Tertiary, RNA, Bacterial genetics, Bacterial Proteins metabolism, Circadian Rhythm Signaling Peptides and Proteins metabolism, Legionella pneumophila genetics, Operon, Stress, Physiological
- Abstract
Legionella pneumophila uses aquatic protozoa as replication niche and protection from harsh environments. Although L. pneumophila is not known to have a circadian clock, it encodes homologues of the KaiBC proteins of Cyanobacteria that regulate circadian gene expression. We show that L. pneumophila kaiB, kaiC and the downstream gene lpp1114, are transcribed as a unit under the control of the stress sigma factor RpoS. KaiC and KaiB of L. pneumophila do not interact as evidenced by yeast and bacterial two-hybrid analyses. Fusion of the C-terminal residues of cyanobacterial KaiB to Legionella KaiB restores their interaction. In contrast, KaiC of L. pneumophila conserved autophosphorylation activity, but KaiB does not trigger the dephosphorylation of KaiC like in Cyanobacteria. The crystal structure of L. pneumophila KaiB suggests that it is an oxidoreductase-like protein with a typical thioredoxin fold. Indeed, mutant analyses revealed that the kai operon-encoded proteins increase fitness of L. pneumophila in competitive environments, and confer higher resistance to oxidative and sodium stress. The phylogenetic analysis indicates that L. pneumophila KaiBC resemble Synechosystis KaiC2B2 and not circadian KaiB1C1. Thus, the L. pneumophila Kai proteins do not encode a circadian clock, but enhance stress resistance and adaption to changes in the environments., (© 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.)
- Published
- 2014
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29. Assessing functional impairment in siblings living with children with disability.
- Author
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Goudie A, Havercamp S, Jamieson B, and Sahr T
- Subjects
- Adaptation, Psychological, Adolescent, Affective Symptoms diagnosis, Child, Child Behavior Disorders diagnosis, Child, Preschool, Early Diagnosis, Female, Health Services Needs and Demand statistics & numerical data, Humans, Interpersonal Relations, Learning Disabilities diagnosis, Learning Disabilities epidemiology, Learning Disabilities psychology, Leisure Activities, Male, Mental Health Services statistics & numerical data, Peer Group, Personality Assessment, Retrospective Studies, Risk Factors, Social Environment, United States, Affective Symptoms epidemiology, Affective Symptoms psychology, Child Behavior Disorders epidemiology, Child Behavior Disorders psychology, Disabled Children psychology, Siblings psychology
- Abstract
Objective: The purpose of this study was to empirically test if siblings of children with disability had higher levels of parent-reported behavioral and emotional functional impairment compared with a peer group of siblings residing with only typically developing children., Methods: This was a retrospective secondary analysis of data from the Medical Expenditure Panel Survey. We included only households with at least 2 children to ensure sibling relationships. Two groups of siblings were formed: 245 siblings resided in households with a child with disability and 6564 siblings resided in households with typically developing children. Parents responded to questions from the Columbia Impairment Scale to identify functional impairment in their children., Results: On the basis of parent reports and after adjusting for sibling demographic characteristics and household background, siblings of children with disability were more likely than siblings residing with typically developing children to have problems with interpersonal relationships, psychopathological functioning, functioning at school, and use of leisure time (P < .05). The percentage of siblings of children with disability classified with significant functional impairment was 16.0% at the first measurement period and 24.2% at the second (P < .001). For siblings of typically developing children there was a smaller percentage increase from 9.5% to 10.3% (P < .001)., Conclusions: Functional impairment is a key indicator for the need of mental health services and, as such, early assessment and interventions to limit increasing severity and short- to long-term consequences need to be addressed. Health care professionals need to consider a family-based health care approach for families raising children with disability.
- Published
- 2013
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30. The Legionella pneumophila orphan sensor kinase LqsT regulates competence and pathogen-host interactions as a component of the LAI-1 circuit.
- Author
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Kessler A, Schell U, Sahr T, Tiaden A, Harrison C, Buchrieser C, and Hilbi H
- Subjects
- Acanthamoeba castellanii microbiology, Bacterial Proteins genetics, Gene Deletion, Gene Expression Regulation, Bacterial, Legionella pneumophila drug effects, Legionella pneumophila genetics, Legionella pneumophila growth & development, Phosphotransferases genetics, Quorum Sensing genetics, Salts pharmacology, Bacterial Proteins metabolism, DNA Transformation Competence genetics, Host-Pathogen Interactions physiology, Legionella pneumophila physiology, Phosphotransferases metabolism, Transcription Factors metabolism
- Abstract
Legionella pneumophila is an amoeba-resistant opportunistic pathogen that performs cell-cell communication through the signalling molecule 3-hydroxypentadecane-4-one (LAI-1, Legionella autoinducer-1). The lqs (Legionella quorum sensing) gene cluster encodes the LAI-1 autoinducer synthase LqsA, the cognate sensor kinase LqsS and the response regulator LqsR. Here we show that the Lqs system includes an 'orphan' homologue of LqsS termed LqsT. Compared with wild-type L. pneumophila, strains lacking lqsT or both lqsS and lqsT show increased salt resistance, greatly enhanced natural competence for DNA acquisition and impaired uptake by phagocytes. Sensitive novel single round growth assays and competition experiments using Acanthamoeba castellanii revealed that ΔlqsT and ΔlqsS-ΔlqsT, as well as ΔlqsA and other lqs mutant strains are impaired for intracellular growth and cannot compete against wild-type bacteria upon co-infection. In contrast to the ΔlqsS strain, ΔlqsT does not produce extracellular filaments. The phenotypes of the ΔlqsS-ΔlqsT strain are partially complemented by either lqsT or lqsS, but are not reversed by overexpression of lqsA, suggesting that LqsT and LqsS are the sole LAI-1-responsive sensor kinases in L. pneumophila. In agreement with the different phenotypes of the ΔlqsT and ΔlqsS strains, lqsT and lqsS are differentially expressed in the post-exponential growth phase, and transcriptome studies indicated that 90% of the genes, which are downregulated in absence of lqsT, are upregulated in absence of lqsS. Reciprocally regulated genes encode components of a 133 kb genomic 'fitness island' or translocated effector proteins implicated in virulence. Together, these results reveal a unique organization of the L. pneumophila Lqs system comprising two partially antagonistic LAI-1-responsive sensor kinases, LqsT and LqsS, which regulate distinct pools of genes implicated in pathogen-host cell interactions, competence, expression of a genomic island or production of extracellular filaments., (© 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.)
- Published
- 2013
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31. Co-immunoprecipitation: protein-RNA and protein-DNA interaction.
- Author
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Sahr T and Buchrieser C
- Subjects
- Chromatin Immunoprecipitation methods, Gene Library, High-Throughput Nucleotide Sequencing methods, Protein Binding, Bacterial Proteins metabolism, DNA, Bacterial metabolism, Immunoprecipitation methods, Legionella pneumophila genetics, Legionella pneumophila metabolism, RNA, Bacterial metabolism
- Abstract
Transcriptional and posttranscriptional regulators play a critical role in allowing a bacterium to adapt to the diverse environments and conditions it encounters. In order to characterize the role of these regulators the identification of their specific interaction partners is of utmost importance. Co-immunoprecipitation (IP) is based on antigen/antibody complex formation to purify a protein of interest from the rest of the samples together with its interaction partner. This method allows us to study direct interaction of a regulator with its specific binding partners like protein-RNA, protein-DNA, or protein-protein interactions. IP typically requires careful optimization and troubleshooting depending on the varying physicochemical characteristics of the protein of interest. In this chapter we present a starting point and the basic guidelines to obtain the best possible results from an IP experiment with subsequent use of new-generation sequencing techniques to detect mRNA or ncRNA targets (RIPseq) and protein-DNA interactions (ChIPseq).
- Published
- 2013
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32. cDNA library construction for next-generation sequencing to determine the transcriptional landscape of Legionella pneumophila.
- Author
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Sahr T and Buchrieser C
- Subjects
- Gene Expression Profiling, RNA, Bacterial isolation & purification, Transcription Initiation Site, Gene Library, High-Throughput Nucleotide Sequencing, Legionella pneumophila genetics, Transcriptome
- Abstract
The adaptation of Legionella pneumophila to the different conditions it encounters in the environment and in the host is governed by a complex regulatory system. Current knowledge of these regulatory networks and the transcriptome responses of L. pneumophila is mainly based on microarray analysis and limited to transcriptional products of annotated protein-coding genes. The application of the Next-Generation Sequencing (NGS) technology allows now genome-wide strand-specific sequencing and accurate determination of all expressed regions of the genome to reveal the complete transcriptional network and the dynamic interplay of specific regulators on a genome-wide level. NGS-based techniques promote deeper understanding of the global transcriptional organization of L. pneumophila by identifying transcription start sites (TSS), alternative TSS and operon organization, noncoding RNAs, antisense RNAs, and 5'-/3'-untranslated regions. In this chapter we describe the construction of cDNA libraries for (1) RNA deep sequencing (RNA-seq) and (2) TSS mapping using the Illumina technology.
- Published
- 2013
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33. Deep sequencing defines the transcriptional map of L. pneumophila and identifies growth phase-dependent regulated ncRNAs implicated in virulence.
- Author
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Sahr T, Rusniok C, Dervins-Ravault D, Sismeiro O, Coppee JY, and Buchrieser C
- Subjects
- 5' Untranslated Regions, Acanthamoeba castellanii microbiology, Bacterial Proteins genetics, Base Sequence, Chromosome Mapping, Consensus Sequence, High-Throughput Nucleotide Sequencing, Legionella pneumophila growth & development, Legionella pneumophila pathogenicity, Molecular Sequence Annotation, Molecular Sequence Data, Nucleic Acid Conformation, Operon, Promoter Regions, Genetic, RNA, Bacterial metabolism, RNA, Small Untranslated metabolism, Sequence Analysis, RNA, Transcription Initiation Site, Transcriptome, Virulence genetics, Gene Expression Regulation, Bacterial, Legionella pneumophila genetics, RNA, Bacterial genetics, RNA, Small Untranslated genetics
- Abstract
The bacterium Legionella pneumophila is found ubiquitously in aquatic environments and can cause a severe pneumonia in humans called Legionnaires' disease. How this bacterium switches from intracellular to extracellular life and adapts to different hosts and environmental conditions is only partly understood. Here we used RNA deep sequencing from exponentially (replicative) and post exponentially (virulent) grown L. pneumophila to analyze the transcriptional landscape of its entire genome. We established the complete operon map and defined 2561 primary transcriptional start sites (TSS). Interestingly, 187 of the 1805 TSS of protein-coding genes contained tandem promoters of which 93 show alternative usage dependent on the growth phase. Similarly, over 60% of 713 here identified ncRNAs are phase dependently regulated. Analysis of their conservation among the seven L. pneumophila genomes sequenced revealed many strain specific differences suggesting that L. pneumophila contains a highly dynamic pool of ncRNAs. Analysis of six ncRNAs exhibiting the same expression pattern as virulence genes showed that two, Lppnc0584 and Lppnc0405 are indeed involved in intracellular growth of L. pneumophila in A. castellanii. Furthermore, L. pneumophila encodes a small RNA named RsmX that functions together with RsmY and RsmZ in the LetA-CsrA regulatory pathway, crucial for the switch to the virulent phenotype. Together our data provide new insight into the transcriptional organization of the L. pneumophila genome, identified many new ncRNAs and will provide a framework for the understanding of virulence and adaptation properties of L. pneumophila.
- Published
- 2012
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34. Legionella pneumophila transcriptional response to chlorine treatment.
- Author
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Bodet C, Sahr T, Dupuy M, Buchrieser C, and Héchard Y
- Subjects
- Bacterial Proteins genetics, Bacterial Proteins metabolism, Gene Expression Profiling, Genes, Bacterial genetics, Legionella pneumophila pathogenicity, Stress, Physiological drug effects, Stress, Physiological genetics, Virulence drug effects, Virulence genetics, Chlorine pharmacology, Gene Expression Regulation, Bacterial drug effects, Legionella pneumophila drug effects, Legionella pneumophila genetics, Transcription, Genetic drug effects
- Abstract
Legionella pneumophila is a ubiquitous environmental microorganism found in freshwater that can cause an acute form of pneumonia known as Legionnaires' disease. Despite widespread use of chlorine to ensure drinking water quality and awareness that L. pneumophila may escape these treatments, little is known about its effects on L. pneumophila. The aim of this study was to investigate the L. pneumophila transcriptional response induced by chlorine treatment. Transcriptome analysis, using DNA arrays, showed that a sublethal dose of chlorine induces a differential expression of 391 genes involved in stress response, virulence, general metabolism, information pathways and transport. Many of the stress response genes were significantly upregulated, whereas a significant number of virulence genes were repressed. In particular, exposure of L. pneumophila to chlorine induced the expression of cellular antioxidant proteins, stress proteins and transcriptional regulators. In addition, glutathione S-transferase specific activity was enhanced following chlorine treatment. Our results clearly indicate that chlorine induces expression of proteins involved in cellular defence mechanisms against oxidative stress that might be involved in adaptation or resistance to chlorine treatment., (Copyright © 2011 Elsevier Ltd. All rights reserved.)
- Published
- 2012
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35. Fatty acid composition modulates sensitivity of Legionella pneumophila to warnericin RK, an antimicrobial peptide.
- Author
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Verdon J, Labanowski J, Sahr T, Ferreira T, Lacombe C, Buchrieser C, Berjeaud JM, and Héchard Y
- Subjects
- Bacterial Proteins pharmacology, Cell Membrane chemistry, Cell Membrane Permeability drug effects, Dose-Response Relationship, Drug, Drug Resistance, Bacterial, Gene Expression Profiling, Gene Expression Regulation, Bacterial, Legionella pneumophila genetics, Membrane Lipids chemistry, Microbial Sensitivity Tests, Oligonucleotide Array Sequence Analysis, Staphylococcus metabolism, Anti-Bacterial Agents pharmacology, Bacteriocins pharmacology, Cell Membrane drug effects, Fatty Acids analysis, Legionella pneumophila drug effects
- Abstract
Warnericin RK is an antimicrobial peptide, produced by a Staphyloccocus warneri strain, described to be specifically active against Legionella, the pathogenic bacteria responsible for Legionnaires' disease. Warnericin RK is an amphiphilic alpha-helical peptide, which possesses a detergent-like mode of action. Two others peptides, δ-hemolysin I and II, produced by the same S. warneri strain, are highly similar to S. aureus δ-hemolysin and also display anti-Legionella activity. It has been recently reported that S. aureus δ-hemolysin activity on vesicles is likewise related to phospholipid acyl-chain structure, such as chain length and saturation. As staphylococcal δ-hemolysins were highly similar, we thus hypothesized that fatty acid composition of Legionella's membrane might influence the sensitivity of the bacteria to warnericin RK. Relationship between sensitivity to the peptide and fatty acid composition was then followed in various conditions. Cells in stationary phase, which were already described as less resistant than cells in exponential phase, displayed higher amounts of branched-chain fatty acids (BCFA) and short chain fatty acids. An adapted strain, able to grow at a concentration 33 fold higher than minimal inhibitory concentration of the wild type (i.e. 1μM), was isolated after repeated transfers of L. pneumophila in the presence of increased concentrations of warnericin RK. The amount of BCFA was significantly higher in the adapted strain than in the wild type strain. Also, a transcriptomic analysis of the wild type and adapted strains showed that two genes involved in fatty acid biosynthesis were repressed in the adapted strain. These genes encode enzymes involved in desaturation and elongation of fatty acids respectively. Their repression was in agreement with the decrease of unsaturated fatty acids and fatty acid chain length in the adapted strain. Conclusively, our results indicate that the increase of BCFA and the decrease of fatty acid chain length in membrane were correlated with the increase in resistance to warnericin RK. Therefore, fatty acid profile seems to play a critical role in the sensitivity of L. pneumophila to warnericin RK., (Copyright © 2010 Elsevier B.V. All rights reserved.)
- Published
- 2011
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36. O-carboxyl- and N-methyltransferases active on plant aquaporins.
- Author
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Sahr T, Adam T, Fizames C, Maurel C, and Santoni V
- Subjects
- Amino Acid Sequence, Aquaporins chemistry, Aquaporins metabolism, Arabidopsis chemistry, Arabidopsis cytology, Arabidopsis enzymology, Arabidopsis genetics, Arabidopsis Proteins chemistry, Arabidopsis Proteins genetics, Biological Transport, Methylation, Molecular Sequence Data, Peptides chemistry, Phylogeny, Plants, Genetically Modified chemistry, Plants, Genetically Modified enzymology, Plants, Genetically Modified genetics, Protein Methyltransferases genetics, Protein O-Methyltransferase genetics, Recombinant Proteins genetics, Recombinant Proteins isolation & purification, Recombinant Proteins metabolism, S-Adenosylmethionine chemistry, Substrate Specificity, Arabidopsis metabolism, Arabidopsis Proteins metabolism, Plants, Genetically Modified metabolism, Protein Methyltransferases metabolism, Protein O-Methyltransferase metabolism
- Abstract
Methylation of biologically active molecules is achieved by methyltransferases (MTases). MTases can act on proteins through N- or O-carboxylmethylation reactions. Methylation of lysine and glutamic acid residues was recently described on the N-terminal tail of AtPIP2;1, a plasma membrane aquaporin of plants. In this study, we combine a bioinformatic and a biochemical screen and identify two MTases of Arabidopsis thaliana, SDG7 (At2g44150) and OMTF3 (At3g61990), as acting on the N-terminal tail of AtPIP2;1, at Lys3 and Glu6, respectively. Confocal microscopy imaging showed the two enzymes to be associated with the endoplasmic reticulum. An in vitro assay using various AtPIP2;1 N-terminal peptides as a bait allowed characterization of the enzymatic properties of recombinant SDG7 and OMTF3. The two enzymes showed minimal apparent K(m) values for their substrates, S-adenosylmethionine and peptide, in the range of 5-8 and 2-9 μM, respectively. SDG7 was shown to almost exclusively mono- or di-methylate Lys3. In contrast, OMTF3 specifically methylated Glu6, this methylation being dependent on the methylation profile of the neighboring Lys3 residue. In conclusion, this study allows the characterization of the first MTases able to methylate plant transmembrane proteins and provides the first identification of a glutamate-MTase in eukaryotes.
- Published
- 2010
- Full Text
- View/download PDF
37. The Legionella pneumophila LetA/LetS two-component system exhibits rheostat-like behavior.
- Author
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Edwards RL, Jules M, Sahr T, Buchrieser C, and Swanson MS
- Subjects
- Amino Acid Substitution genetics, Animals, Bacterial Proteins genetics, Gene Deletion, Macrophages microbiology, Mice, Mutagenesis, Site-Directed, Mutation, Missense, Phosphates metabolism, Bacterial Proteins physiology, Gene Expression Regulation, Bacterial, Legionella pneumophila physiology, Signal Transduction
- Abstract
When confronted with metabolic stress, replicative Legionella pneumophila bacteria convert to resilient, infectious cells equipped for transmission. Differentiation is promoted by the LetA/LetS two-component system, which belongs to a family of signal-transducing proteins that employ a four-step phosphorelay to regulate gene expression. Histidine 307 of LetS was essential to switch on the transmission profile, but a threonine substitution at position 311 (T311M) suggested a rheostat-like function. The letS(T311M) bacteria resembled the wild type (WT) for some traits and letS null mutants for others, whereas they displayed intermediate levels of infectivity, cytotoxicity, and lysosome evasion. Although only 30 to 50% of letS(T311M) mutants became motile, flow cytometry determined that every cell eventually activated the flagellin promoter to WT levels, but expression was delayed. Likewise, letS(T311M) mutants exhibited delayed induction of RsmY and RsmZ, regulatory RNAs that relieve CsrA repression of transmission traits. Transcriptional profile analysis revealed that letS(T311M) mutants expressed the flagellar regulon and multiple other transmissive-phase loci at a higher cell density than the WT. Accordingly, we postulate that the letS(T311M) mutant may relay phosphate less efficiently than the WT LetS sensor protein, leading to sluggish gene expression and a variety of phenotypic profiles. Thus, as first described for BvgA/BvgS, rather than acting as on/off switches, this family of two-component systems exhibit rheostat activity that likely confers versatility as microbes adapt to fluctuating environments.
- Published
- 2010
- Full Text
- View/download PDF
38. The autoinducer synthase LqsA and putative sensor kinase LqsS regulate phagocyte interactions, extracellular filaments and a genomic island of Legionella pneumophila.
- Author
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Tiaden A, Spirig T, Sahr T, Wälti MA, Boucke K, Buchrieser C, and Hilbi H
- Subjects
- Acanthamoeba castellanii growth & development, Acanthamoeba castellanii microbiology, Animals, Bacterial Proteins genetics, Cell Line, Gene Expression Profiling, HL-60 Cells, Histidine Kinase, Host-Pathogen Interactions, Humans, Intermediate Filaments, Legionella pneumophila genetics, Legionella pneumophila metabolism, Legionella pneumophila pathogenicity, Mice, Mutation, Oligonucleotide Array Sequence Analysis, Phagocytes immunology, Protein Kinases genetics, Transcription Factors genetics, Virulence, Bacterial Proteins metabolism, Gene Expression Regulation, Bacterial, Genomic Islands physiology, Legionella pneumophila physiology, Phagocytes microbiology, Protein Kinases metabolism, Transcription Factors metabolism
- Abstract
The amoebae-resistant opportunistic pathogen Legionella pneumophila employs a biphasic life cycle to replicate in host cells and spread to new niches. Upon entering the stationary growth phase, the bacteria switch to a transmissive (virulent) state, which involves a complex regulatory network including the lqs gene cluster (lqsA-lqsR-hdeD-lqsS). LqsR is a putative response regulator that promotes host-pathogen interactions and represses replication. The autoinducer synthase LqsA catalyses the production of the diffusible signalling molecule 3-hydroxypentadecan-4-one (LAI-1) that is presumably recognized by the sensor kinase LqsS. Here, we analysed L. pneumophila strains lacking lqsA or lqsS. Compared with wild-type L. pneumophila, the DeltalqsS strain was more salt-resistant and impaired for the Icm/Dot type IV secretion system-dependent uptake by phagocytes. Legionella pneumophila strains lacking lqsS, lqsR or the alternative sigma factor rpoS sedimented more slowly and produced extracellular filaments. Deletion of lqsA moderately reduced the uptake of L. pneumophila by phagocytes, and the defect was complemented by expressing lqsA in trans. Unexpectedly, the overexpression of lqsA also restored the virulence defect and reduced filament production of L. pneumophila mutant strains lacking lqsS or lqsR, but not the phenotypes of strains lacking rpoS or icmT. These results suggest that LqsA products also signal through sensors not encoded by the lqs gene cluster. A transcriptome analysis of the DeltalqsA and DeltalqsS mutant strains revealed that under the conditions tested, lqsA regulated only few genes, whereas lqsS upregulated the expression of 93 genes at least twofold. These include 52 genes clustered in a 133 kb high plasticity genomic island, which is flanked by putative DNA-mobilizing genes and encodes multiple metal ion efflux pumps. Upon overexpression of lqsA, a cluster of 19 genes in the genomic island was also upregulated, suggesting that LqsA and LqsS participate in the same regulatory circuit.
- Published
- 2010
- Full Text
- View/download PDF
39. Distinct roles of ppGpp and DksA in Legionella pneumophila differentiation.
- Author
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Dalebroux ZD, Yagi BF, Sahr T, Buchrieser C, and Swanson MS
- Subjects
- Amino Acid Sequence, Bacterial Proteins genetics, Cell Adhesion, Cell Survival, Colony Count, Microbial, Flagella physiology, Gene Deletion, Gene Expression Profiling, Legionella pneumophila growth & development, Legionella pneumophila metabolism, Legionella pneumophila pathogenicity, Macrophages microbiology, Molecular Sequence Data, Virulence Factors genetics, Bacterial Proteins metabolism, Gene Expression Regulation, Bacterial, Guanosine Tetraphosphate metabolism, Legionella pneumophila physiology, Virulence Factors metabolism
- Abstract
To transit between hosts, intracellular Legionella pneumophila transform into a motile, infectious, transmissive state. Here we exploit the pathogen's life cycle to examine how guanosine tetraphosphate (ppGpp) and DksA cooperate to govern bacterial differentiation. Transcriptional profiling revealed that during transmission alarmone accumulation increases the mRNA for flagellar and Type IV-secretion components, secreted host effectors and regulators, and decreases transcripts for translation, membrane modification and ATP synthesis machinery. DksA is critical for differentiation, since mutants are defective for stationary phase survival, flagellar gene activation, lysosome avoidance and macrophage cytotoxicity. The roles of ppGpp and DksA depend on the context. For macrophage transmission, ppGpp is essential, whereas DksA is dispensable, indicating that ppGpp can act autonomously. In broth, DksA promotes differentiation when ppGpp levels increase, or during fatty acid stress, as judged by flaA expression and evasion of degradation by macrophages. For flagella morphogenesis, DksA is required for basal fliA (sigma(28)) promoter activity. When alarmone levels increase, DksA cooperates with ppGpp to generate a pulse of Class II rod RNA or to amplify the Class III sigma factor and Class IV flagellin RNAs. Thus, DksA responds to the level of ppGpp and other stress signals to co-ordinate L. pneumophila differentiation.
- Published
- 2010
- Full Text
- View/download PDF
40. Control of flagellar gene regulation in Legionella pneumophila and its relation to growth phase.
- Author
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Albert-Weissenberger C, Sahr T, Sismeiro O, Hacker J, Heuner K, and Buchrieser C
- Subjects
- Animals, Bacterial Proteins genetics, Blotting, Western, Cell Line, Cyclic GMP analogs & derivatives, Cyclic GMP metabolism, Cyclic GMP physiology, Flagella genetics, Flagella ultrastructure, Flagellin genetics, Flagellin metabolism, Legionella pneumophila genetics, Legionella pneumophila ultrastructure, Mice, Microscopy, Electron, Transmission, Mutation, Oligonucleotide Array Sequence Analysis, Polymerase Chain Reaction, Bacterial Proteins physiology, Flagella physiology, Gene Expression Regulation, Bacterial genetics, Gene Expression Regulation, Bacterial physiology, Legionella pneumophila growth & development, Legionella pneumophila metabolism
- Abstract
The bacterial pathogen Legionella pneumophila responds to environmental changes by differentiation. At least two forms are well described: replicative bacteria are avirulent; in contrast, transmissive bacteria express virulence traits and flagella. Phenotypic analysis, Western blotting, and electron microscopy of mutants of the regulatory genes encoding RpoN, FleQ, FleR, and FliA demonstrated that flagellin expression is strongly repressed and that the mutants are nonflagellated in the transmissive phase. Transcriptome analyses elucidated that RpoN, together with FleQ, enhances transcription of 14 out of 31 flagellar class II genes, which code for the basal body, hook, and regulatory proteins. Unexpectedly, FleQ independent of RpoN enhances the transcription of fliA encoding sigma 28. Expression analysis of a fliA mutant showed that FliA activates three out of the five remaining flagellar class III genes and the flagellar class IV genes. Surprisingly, FleR does not induce but inhibits expression of at least 14 flagellar class III genes on the transcriptional level. Thus, we propose that flagellar class II genes are controlled by FleQ and RpoN, whereas the transcription of the class III gene fliA is controlled in a FleQ-dependent but RpoN-independent manner. However, RpoN and FleR might influence flagellin synthesis on a posttranscriptional level. In contrast to the commonly accepted view that enhancer-binding proteins such as FleQ always interact with RpoN to fullfill their regulatory functions, our results strongly indicate that FleQ regulates gene expression that is RpoN dependent and RpoN independent. Finally, FliA induces expression of flagellar class III and IV genes leading to the complete synthesis of the flagellum.
- Published
- 2010
- Full Text
- View/download PDF
41. Two small ncRNAs jointly govern virulence and transmission in Legionella pneumophila.
- Author
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Sahr T, Brüggemann H, Jules M, Lomma M, Albert-Weissenberger C, Cazalet C, and Buchrieser C
- Subjects
- Bacterial Proteins genetics, Base Sequence, Cell Line, Gene Expression Profiling, Gene Expression Regulation, Bacterial, Humans, Legionella pneumophila genetics, Molecular Sequence Data, RNA, Bacterial genetics, RNA, Untranslated genetics, Bacterial Proteins metabolism, Legionella pneumophila pathogenicity, RNA, Bacterial metabolism, RNA, Untranslated metabolism, Virulence
- Abstract
To transit from intra- to extracellular environments, Legionella pneumophila differentiates from a replicative/non-virulent to a transmissive/virulent form using the two-component system LetA/LetS and the global repressor protein CsrA. While investigating how both regulators act co-ordinately we characterized two ncRNAs, RsmY and RsmZ, that link the LetA/LetS and CsrA regulatory networks. We demonstrate that LetA directly regulates their expression and show that RsmY and RsmZ are functional in Escherichia coli and are able to bind CsrA in vitro. Single mutants have no (ΔrsmY) or a little (ΔrsmZ) impact on virulence, but the ΔrsmYZ strain shows a drastic defect in intracellular growth in Acanthamoeba castellanii and THP-1 monocyte-derived macrophages. Analysis of the transcriptional programmes of the ΔletA, ΔletS and ΔrsmYZ strains revealed that the switch to the transmissive phase is partially blocked. One major difference between the ΔletA, ΔletS and ΔrsmYZ strains was that the latter synthesizes flagella. Taken together, LetA activates transcription of RsmY and RsmZ, which sequester CsrA and abolish its post-transcriptional repressive activity. However, the RsmYZ-CsrA pathway appears not to be the main or only regulatory circuit governing flagella synthesis. We suggest that rather RpoS and LetA, by influencing LetE and probably cyclic-di-GMP levels, regulate motility in L. pneumophila.
- Published
- 2009
- Full Text
- View/download PDF
42. The dose-response relationship of adolescent religious activity and substance use: variation across demographic groups.
- Author
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Steinman KJ, Ferketich AK, and Sahr T
- Subjects
- Adolescent, Demography, Family Characteristics, Female, Humans, Juvenile Delinquency prevention & control, Male, Models, Theoretical, Ohio epidemiology, Regression Analysis, Substance-Related Disorders epidemiology, Substance-Related Disorders ethnology, Surveys and Questionnaires, Adolescent Behavior, Religion, Substance-Related Disorders prevention & control
- Abstract
This article addresses two inconsistent findings in the literature on adolescent religious activity (RA) and substance use: whether a dose-response relationship characterizes the association of these variables, and whether the association varies by grade, gender, ethnicity, family structure, school type, and type of substance. Multinomial logistic regression analyses of a large, diverse data set of high school students in metropolitan Columbus, Ohio ( n = 33,007), found marked differences in alcohol, marijuana, and cigarette use among youths who never, occasionally, or regularly participated in RA. Weekly RA was consistently associated with less substance use, yet occasional RA sometimes was associated with greater use. Four groups accounted for variations in the RA-substance use relationship: African American youths, younger White youths, 12th-grade White males, and 12th-grade White females. Researchers should avoid assuming the RA-substance use relationship is dose-response and consider the implications of this complexity for theory and practice.
- Published
- 2008
- Full Text
- View/download PDF
43. Folate synthesis in plants: purification, kinetic properties, and inhibition of aminodeoxychorismate synthase.
- Author
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Sahr T, Ravanel S, Basset G, Nichols BP, Hanson AD, and Rébeillé F
- Subjects
- 4-Aminobenzoic Acid metabolism, Arabidopsis Proteins antagonists & inhibitors, Arabidopsis Proteins genetics, Arabidopsis Proteins metabolism, Carbon-Nitrogen Ligases antagonists & inhibitors, Carbon-Nitrogen Ligases genetics, Carbon-Nitrogen Ligases metabolism, Chorismic Acid metabolism, Escherichia coli, Folic Acid analogs & derivatives, Folic Acid pharmacology, Glutamine metabolism, Kinetics, Methotrexate pharmacology, Pyruvic Acid metabolism, Recombinant Fusion Proteins antagonists & inhibitors, Recombinant Fusion Proteins metabolism, Substrate Specificity, Transaminases, Arabidopsis Proteins isolation & purification, Carbon-Nitrogen Ligases isolation & purification
- Abstract
pABA (p-aminobenzoate) is a precursor of folates and, besides esterification to glucose, has no other known metabolic fate in plants. It is synthesized in two steps from chorismate and glutamine, the first step being their conversion into glutamate and ADC (4-aminodeoxychorismate). In Escherichia coli, two proteins forming a heterodimeric complex are required for this reaction, but, in plants and lower eukaryotes, a single protein is involved. The Arabidopsis enzyme was expressed in E. coli and was purified to homogeneity. The monomeric enzyme (95 kDa) catalyses two reactions: release of NH3 from glutamine (glutaminase activity) and substitution of NH3 for the hydroxy group at position 4 of chorismate (ADC synthase activity). The kinetic parameters of the plant enzyme are broadly similar to those of the bacterial complex, with K(m) values for glutamine and chorismate of 600 and 1.5 microM respectively. As with the bacterial enzyme, externally added NH3 was a very poor substrate for the plant enzyme, suggesting that NH3 released from glutamine is preferentially channelled to chorismate. The glutaminase activity could operate alone, but the presence of chorismate increased the efficiency of the reaction 10-fold, showing the interdependency of the two domains. The plant enzyme was inhibited by dihydrofolate and its analogue methotrexate, a feature never reported for the prokaryotic system. These molecules were inhibitors of the glutaminase reaction, competitive with respect to glutamine (K(i) values of 10 and 1 microM for dihydrofolate and methotrexate respectively). These findings support the view that the monomeric ADC synthase is a potential target for antifolate drugs.
- Published
- 2006
- Full Text
- View/download PDF
44. Differences in the kinetics and scale of signalling molecule production modulate the ozone sensitivity of hybrid poplar clones: the roles of H2O2, ethylene and salicylic acid.
- Author
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Diara C, Castagna A, Baldan B, Sodi AM, Sahr T, Langebartels C, Sebastiani L, and Ranieri A
- Subjects
- Base Sequence, DNA, Plant genetics, Ethylenes metabolism, Gene Expression drug effects, Genes, Plant drug effects, Hydrogen Peroxide metabolism, Kinetics, Lyases genetics, Populus genetics, Salicylic Acid metabolism, Signal Transduction drug effects, Ozone toxicity, Populus drug effects, Populus metabolism
- Abstract
Hydrogen peroxide (H(2)O(2)), ethylene, 1-aminocyclopropane-1-carboxylic acid (ACC) and salicylic acid (SA) concentrations and ACC synthase (ACS) gene expression were measured to establish whether the high sensitivity of the Populus deltoides x maximowiczii clone Eridano to ozone (O(3)) exposure, compared with the O(3)-resistant Populus deltoides x euramericana clone I-214, is attributable to differences in the modulation of signal transduction pathways. In a time-course experiment, Populus deltoides (poplar) clones were exposed to acute fumigation with 150 nl l(-1) O(3) for 5 h. The two poplar clones showed differences in ethylene evolution, I-214 displaying earlier and less pronounced ethylene emission than Eridano. In both clones, ethylene evolution was accompanied by increased ACS transcript levels and enhanced emission of free ACC. I-214 exhibited a greater basal concentration of free SA and a lower concentration of the conjugated pool. However, a slight accumulation of free SA at the end of the 5-h exposure was found only in Eridano, together with an earlier minimal increase in the concentration of conjugated SA. The results show that both clones react to O(3) by producing H(2)O(2), ethylene and SA, but the difference in sensitivity to the pollutant is probably attributable to differences in the kinetics and magnitude of this response.
- Published
- 2005
- Full Text
- View/download PDF
45. Low-level radiocaesium exposure alters gene expression in roots of Arabidopsis.
- Author
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Sahr T, Voigt G, Schimmack W, Paretzke HG, and Ernst D
- Subjects
- Cesium Radioisotopes metabolism, Gene Expression Profiling, Soil Pollutants, Radioactive, Arabidopsis metabolism, Arabidopsis radiation effects, Cesium Radioisotopes toxicity, Gene Expression Regulation, Plant radiation effects
- Abstract
Radiocaesium is one of the main anthropogenic sources of internal and external exposure to beta- and gamma-radiation (e.g. from global fallout of atmospheric atomic bomb testing and from the Chernobyl reactor accident). Here we investigated gene expression by suppression subtractive hybridization (SSH) and reverse transcription-polymerase chain reaction (RT-PCR) in Arabidopsis thaliana, which was induced by the root uptake of 134Cs. SSH analysis resulted in the isolation of 46 clones that were differentially expressed at 30 Bq cm(-3) 134Cs. Most of the expressed sequence tags identified belonged to genes encoding proteins that were involved in cell growth, cell division and the development of plants, and in proteins controlling translation, general metabolism and stress defence, including a DNA excision repair protein. The accumulation of caesium in plant material was measured in plants grown for 5 wk on agar contaminated by up to 60 Bq cm(-3) 134Cs. 134Cs was found to accumulate, in particular, in leaf rosettes and was dependent on the activity concentration in the growth media. The data indicate that low-level ionizing radiation influences important cellular responses, resulting in a changed gene-expression profile.
- Published
- 2005
- Full Text
- View/download PDF
46. Caesium-affected gene expression in Arabidopsis thaliana.
- Author
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Sahr T, Voigt G, Paretzke HG, Schramel P, and Ernst D
- Subjects
- Arabidopsis drug effects, Arabidopsis genetics, Cesium pharmacology, Microarray Analysis, Plant Leaves drug effects, Arabidopsis metabolism, Cesium metabolism, Gene Expression Regulation, Plant drug effects
- Abstract
* Excessive caesium can be toxic to plants. Here we investigated Cs uptake and caesium-induced gene expression in Arabidopsis thaliana. * Accumulation was measured in plants grown for 5 wk on agar supplemented with nontoxic and up to toxic levels of Cs. Caesium-induced gene expression was studied by suppression-subtractive hybridization (SSH) and RT-PCR. * Caesium accumulated in leaf rosettes dependent upon the external concentration in the growth media, whereas the potassium concentration decreased in rosettes. At a concentration of 850 microM, Cs plants showed reduced development, and withered with an increase in concentration to 1 mM Cs. SSH resulted in the isolation of 73 clones that were differentially expressed at a Cs concentration of 150 microM. Most of the genes identified belong to groups of genes encoding proteins in stress defence, detoxification, transport, homeostasis and general metabolism, and proteins controlling transcription and translation. * The present study identified a number of marker genes for Cs in Arabidopsis grown under nontoxic Cs concentrations, indicating that Cs acts as an abiotic stress factor.
- Published
- 2005
- Full Text
- View/download PDF
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