111 results on '"Garbi, C"'
Search Results
2. Increased hexosamine biosynthetic pathway flux dedifferentiates INS-1E cells and murine islets by an extracellular signal-regulated kinase (ERK)1/2-mediated signal transmission pathway
- Author
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Lombardi, A., Ulianich, L., Treglia, A. S., Nigro, C., Parrillo, L., Lofrumento, D. D., Nicolardi, G., Garbi, C., Beguinot, F., Miele, C., and Di Jeso, B.
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- 2012
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3. THYROID CANCER SUSCEPTIBILITY GENE FOXE1 PROMOTES CELL MOTILITY AND IS REQUIRED FOR SURVIVAL OF ADULT THYROID CELLS: OP71
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De Menna, M, Marotta, P, Esposito, R, Orlacchio, A, De Vita, G, Garbi, C, De Felice, M, and Lauro, R Di
- Published
- 2013
4. Propachlor removal by Pseudomonas strain GCH1 in an immobilized-cell system
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Martin, M., Mengs, G., Plaza, E., Garbi, C., Sanchez, M., Gibello, A., Gutierrez, F., and Ferrer, E.
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Bioremediation -- Research ,Herbicides -- Research ,Pseudomonas -- Research ,Biological sciences - Abstract
Researchers describe a bioremediation system containing immobilized cells of Pseudomonas strain GCH1 adsorbed onto a ceramic support column. The cells removed 98% of the herbicide propachlor present in the system.
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- 2000
5. Counterregulation of cAMP-directed kinase activities controls ciliogenesis
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Porpora M, SAUCHELLA, SIMONA, Rinaldi L, Delle Donne R, Sepe M, Torres-Quesada O, Intartaglia D, Garbi C, Insabato L, Santoriello M, Bachmann VA, Synofzik M, Lindner HH, Conte I, Stefan E, Feliciello A., CONTE, IVAN, Porpora, M, Sauchella, Simona, Rinaldi, L, Delle Donne, R, Sepe, M, Torres-Quesada, O, Intartaglia, D, Garbi, C, Insabato, L, Santoriello, M, Bachmann, Va, Synofzik, M, Lindner, Hh, Conte, I, Stefan, E, Feliciello, A., and Conte, Ivan
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PKA, cAMP, proteasome, primary cilium, NEK10, ubiquitin - Abstract
The primary cilium emanates from the cell surface of growth-arrested cells and plays a central role in vertebrate development and tissue homeostasis. The mechanisms that control ciliogenesis have been extensively explored. However, the intersection between GPCR signaling and the ubiquitin pathway in the control of cilium stability is unknown. Here, we observe that cAMP elevation promotes cilia resorption. At centriolar satellites, we identify a multimeric complex nucleated by PCM1 that includes two kinases, NEK10 and PKA, and the E3 ubiquitin ligase CHIP. We show that NEK10 is essential for ciliogenesis in mammals and for the development of medaka fish. PKA phosphorylation primes NEK10 for CHIP-mediated ubiquitination and proteolysis resulting in cilia resorption. Dearangement of this control mechanism occurs in proliferative and genetic disorders. These findings unveil a pericentriolar kinase signalosome that efficiently links the cAMP cascade with the ubiquitin-proteasome system, controlling essential aspects of ciliogenesis.
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- 2018
6. Mitochondrial AKAP1 supports mTOR pathway and tumor growth
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Rinaldi L., Sepe M., Delle Donne R., Conte K., Arcella A., Borzacchiello D., Amente S., De Vita F., Porpora M., Garbi C., Oliva M. A., Procaccini C., Faicchia D., Matarese G., Zito Marino F., Rocco G., Pignatiello S., Franco R., Insabato L., Majello B., Feliciello A., ZITO MARINO, Federica, Rinaldi, L., Sepe, M., Delle Donne, R., Conte, K., Arcella, A., Borzacchiello, D., Amente, S., De Vita, F., Porpora, M., Garbi, C., Oliva, M. A., Procaccini, C., Faicchia, D., Matarese, G., Zito Marino, F., Rocco, G., Pignatiello, S., Franco, R., Insabato, L., Majello, B., Feliciello, A., ZITO MARINO, Federica, Rinaldi, Laura, Sepe, Maria, Delle Donne, Rossella, Conte, Kristel, Arcella, Antonietta, Borzacchiello, Domenica, Amente, Stefano, De Vita, Fernanda, Porpora, Monia, Garbi, Corrado, Oliva, Maria A, Procaccini, Claudio, Faicchia, Deriggio, Matarese, Giuseppe, Zito Marino, Federica, Rocco, Gaetano, Pignatiello, Sara, Franco, Renato, Insabato, Luigi, Majello, Barbara, and Feliciello, Antonio
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0301 basic medicine ,Scaffold protein ,Male ,Cancer Research ,Lung Neoplasms ,Transcription, Genetic ,A Kinase Anchor Proteins ,Mitochondrion ,mTORC2 ,RNA, Small Interfering ,Nuclear Protein ,Mitochondrial ,AKAP1 ,mTOR ,TOR Serine-Threonine Kinase ,Brain Neoplasms ,TOR Serine-Threonine Kinases ,Nuclear Proteins ,A Kinase Anchor Protein ,3. Good health ,Cell biology ,Mitochondria ,Gene Expression Regulation, Neoplastic ,Original Article ,Survival Analysi ,Signal transduction ,Neuroglia ,Protein Binding ,Signal Transduction ,Immunology ,Mice, Nude ,Biology ,Brain Neoplasm ,Proto-Oncogene Proteins c-myc ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,Cell Line, Tumor ,Animals ,PI3K/AKT/mTOR pathway ,Cell Proliferation ,Epithelial Cell ,Cell growth ,Animal ,RPTOR ,Epithelial Cells ,Cell Biology ,Survival Analysis ,Lung Neoplasm ,030104 developmental biology ,Cancer research ,Organelle biogenesis ,Neoplasm Transplantation - Abstract
Mitochondria are the powerhouses of energy production and the sites where metabolic pathway and survival signals integrate and focus, promoting adaptive responses to hormone stimulation and nutrient availability. Increasing evidence suggests that mitochondrial bioenergetics, metabolism and signaling are linked to tumorigenesis. AKAP1 scaffolding protein integrates cAMP and src signaling on mitochondria, regulating organelle biogenesis, oxidative metabolism and cell survival. Here, we provide evidence that AKAP1 is a transcriptional target of Myc and supports the growth of cancer cells. We identify Sestrin2, a leucine sensor and inhibitor of the mammalian target of rapamycin (mTOR), as a novel component of the complex assembled by AKAP1 on mitochondria. Downregulation of AKAP1 impaired mTOR pathway and inhibited glioblastoma growth. Both effects were reversed by concomitant depletion of AKAP1 and sestrin2. High levels of AKAP1 were found in a wide variety of high-grade cancer tissues. In lung cancer, AKAP1 expression correlates with high levels of Myc, mTOR phosphorylation and reduced patient survival. Collectively, these data disclose a previously unrecognized role of AKAP1 in mTOR pathway regulation and cancer growth. AKAP1/mTOR signal integration on mitochondria may provide a new target for cancer therapy.
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- 2017
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7. Application of cyanobacteria oligonucleotide probes designed for identifying bacterial cells in surface water
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Barra Caracciolo A, Dejana L, Fajardo C, Garbi C, Grenni P, Martin M., Mengs G, and Medlin L
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FISH ,algal bloom ,Microcystis aeruginosa ,Planktothrix agardii - Abstract
Cyanobacteria colonize different environments and blooms can occur both in contaminated and noncontaminated water bodies (freshwater, brackish and marine areas). In some cases, some bacterial species can produce toxins and this phenomenon can have a negative impact on ecosystem and human health. Inside 150 genera of cyanobacteria known, more than 40 species are able to produce toxins. The latter are natural compounds which differ from both chemical and toxicological point of view; they are responsible for both acute and chronic poisoning in animals and humans. Among the main classes of cyanotoxins, microcystins are frequently found in the environment. These toxins are mainly accumulated in liver, but also in intestine and kidneys and can be very dangerous for animal and human health [1]. Fast and sensitive methods to identify unequivocally species belonging to Microcystis and Planktothrix genera are necessary to discriminate these genera respect to the other non-toxic cyanobacteria. For this purpose, we designed, developed and validated some oligonucleotide probes for FISH (Fluorescence In Situ Hybridization) analysis in order to detect them in freshwater. The FISH probes were designed using the ARB software with the Silva database in the framework of the MicroCoKit project. We have focused on identification of Microcystis aeruginosa and Planktothrix agardii species because they are among the most common toxic cyanobacteria in freshwater. We tested different fixative methods to minimise the natural autofluorescence from chlorophyll-a and some accessories pigments (e.g. phycobilins and carotenoids) in order to visualize Microcystis aeruginosa and Planktothrix agardii under a laser confocal microscope. Firstly, the FISH probes designed (GNPlankS02, PkAgD03, MicAerD03) have been tested on pure cultures of M. aeruginosa and P. agardii species. Then the probes were successfully applied to several water samples collected from the River Tiber and Lake Albano (Italy).
- Published
- 2017
8. The Type and the Localization of cAMP-dependent Protein Kinase Regulate Transmission of cAMP Signals to the Nucleus in Cortical and Cerebellar Granule Cells
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Paolillo M, FELICIELLO, ANTONIO, PORCELLINI, ANTONIO, Garbi C, Bifulco M, Schinelli S, Ventra C, Ricciardelli G, Schettini G, Avvedimento EV, STABILE, EUGENIO, Paolillo, M, Feliciello, Antonio, Porcellini, Antonio, Garbi, Corrado, Bifulco, M, Schinelli, S, Ventra, C, Stabile, E, Ricciardelli, G, Schettini, G, Avvedimento, VITTORIO ENRICO, Garbi, C, Stabile, Eugenio, and Avvedimento, Ev
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Gene isoform ,CAMP Responsive Element Binding Protein ,Cerebellum ,Protein subunit ,A Kinase Anchor Proteins ,Biology ,Biochemistry ,AKAP ,cAMP ,Cyclic AMP ,medicine ,Animals ,Protein Isoforms ,PKA ,Protein kinase A ,Molecular Biology ,Adaptor Proteins, Signal Transducing ,Cell Nucleus ,Cerebral Cortex ,Neurons ,Proteins ,Cell Biology ,Cyclic AMP-Dependent Protein Kinases ,Molecular biology ,neuron ,Rats ,Cell biology ,medicine.anatomical_structure ,biology.protein ,Carrier Proteins ,CREB1 ,Nucleus ,Signal Transduction - Abstract
cAMP signals are received and transmitted by multiple isoforms of cAMP-dependent protein kinases, typically determined by their specific regulatory subunits. In the brain the major regulatory isoform RIIbeta and the RII-anchor protein, AKAP150 (rat) or 75 (bovine), are differentially expressed. Cortical neurons express RIIbeta and AKAP75; conversely, granule cerebellar cells express predominantly RIalpha and RIIalpha. Cortical neurons accumulate PKA catalytic subunit and phosphorylated cAMP responsive element binding protein very efficiently into nuclei upon cAMP induction, whereas granule cerebellar cells fail to do so. Down-regulation of RIIbeta synthesis by antisense oligonucleotides inhibited cAMP-induced nuclear signaling in cortical neurons. Expression in cerebellar granule cells of RIIbeta and AKAP75 genes by microinjection of specific expression vectors, markedly stimulated cAMP-induced transcription of the lacZ gene driven by a cAMP-responsive element promoter. These data indicate that the composition of PKA in cortical and granule cells underlies the differential ability of these cells to transmit cAMP signals to the nucleus.
- Published
- 1999
9. praja2 regulates KSR1 stability and mitogenic signaling
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Rinaldi, L, primary, Delle Donne, R, additional, Sepe, M, additional, Porpora, M, additional, Garbi, C, additional, Chiuso, F, additional, Gallo, A, additional, Parisi, S, additional, Russo, L, additional, Bachmann, V, additional, Huber, R G, additional, Stefan, E, additional, Russo, T, additional, and Feliciello, A, additional
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- 2016
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10. Proyecto de extensión universitaria: Manipulación de Alimentos, Educación y Seguridad Alimentaria (MESA) relevamiento bromatológico y control de alimentos en instituciones educativas y recreativas de la ciudad de La Plata
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Coll Cárdenas, Fernanda Josefina, Villat, María Cecilia, Copes, Julio Alberto, Garbi, C. J., Pena, Irene del Carmen, Olivera, D., Noia, Miguel Ángel, Laporte, Gladys M., Sota, Pablo Elías de la, Olaiz, Daniel Adrián, Álvarez, María Cecilia, and Rodríguez, V.
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Alimentos ,Ciencias Veterinarias ,Bromatología ,Extensión universitaria - Abstract
El objetivo de este Proyecto es el de formar Agentes Multiplicadores con criterio bromatológico que fomenten normas de control sanitario, aseguren la calidad e inocuidad de los alimentos que llegan al consumidor y apoyen la transformación, a través de Capacitación y Asistencia técnica, teniendo como punto de partida la Educación para la Salud y la Educación Ecológica., Mesa: Alimentos y Vacunas., Facultad de Ciencias Veterinarias
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- 2008
11. La rabia urbana en la provincia de Buenos Aires, Argentina: origen-evolución-actualidad
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Amasino, Carlos Francisco, Garbi, C. J., and Amasino, M. F.
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Ciencias Veterinarias ,Virosis ,Buenos Aires (Argentina) ,rabia ,historia de la rabia ,Buenos Aires ,La Plata ,Instituto Biológico ,Salud Ambiental ,rabies ,rage ,history of the rage ,Buenos Aires province ,Biological Institute ,Rabia - Abstract
La rabia es una enfermedad infecciosa, zoonótica, caracterizada por alteraciones del sistema nervioso, producida por el virus de la rabia, perteneciente al Género Lyssavirus de la Familia Rhabdoviridae. Es habitualmente transmitida por la mordedura de un animal rabioso que elimina el virus por la saliva. La rabia urbana es la que ocurre en las ciudades, siendo el perro el transmisor y afectado más importante, seguido por el gato y el hombre. El presente trabajo presenta un enfoque global sobre la rabia urbana, enfermedad de los animales y el hombre, en el territorio de la provincia de Buenos Aires, Argentina, hasta el año 2001. Esta provincia presentó el mayor foco de rabia urbana que registró el país desde su aparición en 1806 a consecuencia de las actividades de la 1º invasión inglesa, hasta su erradicación en el año 1984. Durante 1976, el territorio provincial registró 4.759 casos de rabia animal y trece decesos humanos. Se efectúa un encuadre de las situaciones geográficas y políticas de las épocas en que se produjeron acontecimientos importantes y del estado de los conocimientos científicos sobre la enfermedad en esos momentos, que condicionaron y caracterizaron la presentación, manejo y control de esta enfermedad en la provincia., Rabies is an infectious and zoonotic disease characterized by alterations of the nervous system, produced by the rabies virus, which belongs to the genus Lyssavirus of the Rhabdoviridae family. The disease is habitually transmitted by the bite of an infected animal, which eliminates the virus in its saliva. Urban rabies is the one that occurs in the cities, being the dog the most important affected and transmitter of the disease, followed by cat and man. The present work shows a global focus on rabies, animal and man affecting disease, in the territory of the province of Buenos Aires, Argentina. This province presented the biggest focus of urban rabies ever registered in the Argentinian Republic from the moment of the first appearance of the disease, consigned in 1806 and due to the activities of the 1 English Invasion, to the moment of the eradication of the disease in 1984. The subsequent panorama, until 2001 is also following presented. During 1976, the province´s territory registered 4759 cases of animal rabies and 13 human deaths. An alignment of the geographical and political situations of the periods during which important events happened is made. Besides, the state of the scientific knowledge about the disease during these periods, which conditioned and characterized the presentation, handling and control of this disease in the province of Buenos Aires, Argentina, is also analyzed., Facultad de Ciencias Veterinarias
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- 2002
12. PED/PEA-15 controls fibroblast motility and wound closure by ERK1/2-dependent mechanisms
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Buonomo, R., Giacco, F., Vasaturo, A., Caserta, S., Guido, S., Pagliara, V., Garbi, C., Mansueto, G., Cassese, A., Perruolo, G., Oriente, F., Miele, C., Beguinot, F., Formisano, P., Buonomo, R., Giacco, F., Vasaturo, A., Caserta, S., Guido, S., Pagliara, V., Garbi, C., Mansueto, G., Cassese, A., Perruolo, G., Oriente, F., Miele, C., Beguinot, F., and Formisano, P.
- Abstract
Item does not contain fulltext, Cell migration is dependent on the control of signaling events that play significant roles in creating contractile force and in contributing to wound closure. We evaluated wound closure in fibroblasts from mice overexpressing (TgPED) or lacking ped/pea-15 (KO), a gene overexpressed in patients with type 2 diabetes. Cultured skin fibroblasts isolated from TgPED mice showed a significant reduction in the ability to recolonize wounded area during scratch assay, compared to control fibroblasts. This difference was observed both in the absence and in the presence of mytomicin C, an inhibitor of mitosis. In time-lapse experiments, TgPED fibroblasts displayed about twofold lower velocity and diffusion coefficient, as compared to controls. These changes were accompanied by reduced spreading and decreased formation of stress fibers and focal adhesion plaques. At the molecular level, TgPED fibroblasts displayed decreased RhoA activation and increased abundance of phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2). Inhibition of ERK1/2 activity by PD98059 restored RhoA activation, cytoskeleton organization and cell motility, and almost completely rescued wound closure of TgPED fibroblasts. Interestingly, skin fibroblasts isolated from KO mice displayed an increased wound closure ability. In vivo, healing of dorsal wounds was delayed in TgPED and accelerated in KO mice. Thus, PED/PEA-15 may affect fibroblast motility by a mechanism, at least in part, mediated by ERK1/2.
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- 2012
13. A new mathematical model to evaluate simazine removal in three different immobilized-biomass reactors
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Martín, M., Casasús, L., Garbi, C., Nande, M., Vargas, R., Robla, J. I., Sánchez, M., Allende, J. L., Martín, M., Casasús, L., Garbi, C., Nande, M., Vargas, R., Robla, J. I., Sánchez, M., and Allende, J. L.
- Abstract
A new mathematical model based on the cinetical Langmuir equation is developed to interpret and predict the effectiveness of simazine (SZ) removal in immobilized-biomass reactor (IBR), to consider herbicide-support affinity (Cx), and herbicide-cell affinity (Cy). Three solid supports: sepiolite monolith, granular sepiolite, and alginate were used in pilot-scale reactors that were inoculated with Klebsiella planticola DSZ. The abiotic process was analysed by measuring the SZ sorption capacity of the reactor supports. Sepiolite monolith showed the maximum value for herbicide-support affinity (28.02±0.9%). The effectiveness of the biotic process was estimated considering the formation of biomass and SZ biodegradation. Granular sepiolite showed either higher affinity with SZ and viability rate (0.90) throughout the process, and SZ removal rate was 3.39±0.06 mg/h. The mathematical model presented in this paper provides useful insights into the interpretation of experimental data as well as prediction for the implementation of biological reactors. © 2007 Elsevier Ltd. All rights reserved.
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- 2008
14. Wnt4 inhibits cell motility induced by oncogenic Ras
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De Menna, M, primary, D'Amato, V, additional, Ferraro, A, additional, Fusco, A, additional, Di Lauro, R, additional, Garbi, C, additional, and De Vita, G, additional
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- 2012
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15. Increased hexosamine biosynthetic pathway flux dedifferentiates INS-1E cells and murine islets by an extracellular signal-regulated kinase (ERK)1/2-mediated signal transmission pathway
- Author
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Lombardi, A., primary, Ulianich, L., additional, Treglia, A. S., additional, Nigro, C., additional, Parrillo, L., additional, Lofrumento, D. D., additional, Nicolardi, G., additional, Garbi, C., additional, Beguinot, F., additional, Miele, C., additional, and Di Jeso, B., additional
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- 2011
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16. TRAP1 and the proteasome regulatory particle TBP7/Rpt3 interact in the endoplasmic reticulum and control cellular ubiquitination of specific mitochondrial proteins
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Amoroso, M R, primary, Matassa, D S, additional, Laudiero, G, additional, Egorova, A V, additional, Polishchuk, R S, additional, Maddalena, F, additional, Piscazzi, A, additional, Paladino, S, additional, Sarnataro, D, additional, Garbi, C, additional, Landriscina, M, additional, and Esposito, F, additional
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- 2011
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17. The tissue-specific pathway regulation cell proliferation are inherited independently in somatic hybrid between thyroid and liver cells
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Veneziani, B. M., Villone, G., Romano, R., DI CARLO, Angelina, Garbi, C., and Tramontano, D.
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- 1991
18. Metabloic effects of adenosine derivate 6N-isopentenyl adenosine in FRTL-5 thyroid cells
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Bifulco, S., Laezza, C., Tedesco, I., Perrillo, R., Romano, A., DI CARLO, Angelina, and Garbi, C.
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- 1991
19. Metabolic effects of N-6-isopentenyl adenosine in FRTL-5 thyroid cells
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Laezza, C., Tedesco, I., Perrillo, B., Romano, A., DI CARLO, Angelina, Garbi, C., and Aloj, S. M.
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- 1991
20. Immobilized Native Bacteria as a Tool for Bioremediation of Soils and Waters: Implementation and Modeling
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Lobo, C., primary, Sanchez, M., additional, Garbi, C., additional, Ferrer, E., additional, Martinez-Iñigo, M.J., additional, Allende, J.L., additional, Martinez, C., additional, Casasas, L., additional, Alonso, R., additional, Gibello, A., additional, and Martin, M., additional
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- 2002
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21. The v-Ki-Ras Oncogene Alters cAMP Nuclear Signaling by Regulating the Location and the Expression of cAMP-dependent Protein Kinase IIβ
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Feliciello, A., primary, Giuliano, P., additional, Porcellini, A., additional, Garbi, C., additional, Obici, S., additional, Mele, E., additional, Angotti, E., additional, Grieco, D., additional, Amabile, G., additional, Cassano, S., additional, Li, Y., additional, Musti, Anna M., additional, Rubin, Charles S., additional, Gottesman, Max E., additional, and Avvedimento, Enrico V., additional
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- 1996
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22. v-ras and protein kinase C dedifferentiate thyroid cells by down-regulating nuclear cAMP-dependent protein kinase A.
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Gallo, A, primary, Benusiglio, E, additional, Bonapace, I M, additional, Feliciello, A, additional, Cassano, S, additional, Garbi, C, additional, Musti, A M, additional, Gottesman, M E, additional, and Avvedimento, E V, additional
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- 1992
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23. TRAP1 and the proteasome regulatory particle TBP7/Rpt3 interact in the endoplasmic reticulum and control cellular ubiquitination of specific mitochondrial proteins.
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Amoroso, M R, Matassa, D S, Laudiero, G, Egorova, A V, Polishchuk, R S, Maddalena, F, Piscazzi, A, Paladino, S, Sarnataro, D, Garbi, C, Landriscina, M, and Esposito, F
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TUMOR necrosis factor receptors ,HEAT shock proteins ,PROTEINS ,MITOCHONDRIA ,ENDOPLASMIC reticulum ,PROTEASOMES - Abstract
Tumor necrosis factor receptor-associated protein-1 (TRAP1) is a mitochondrial (MITO) antiapoptotic heat-shock protein. The information available on the TRAP1 pathway describes just a few well-characterized functions of this protein in mitochondria. However, our group's use of mass-spectrometric analysis identified TBP7, an AAA-ATPase of the 19S proteasomal subunit, as a putative TRAP1-interacting protein. Surprisingly, TRAP1 and TBP7 colocalize in the endoplasmic reticulum (ER), as demonstrated by biochemical and confocal/electron microscopic analyses, and interact directly, as confirmed by fluorescence resonance energy transfer analysis. This is the first demonstration of TRAP1's presence in this cellular compartment. TRAP1 silencing by short-hairpin RNAs, in cells exposed to thapsigargin-induced ER stress, correlates with upregulation of BiP/Grp78, thus suggesting a role of TRAP1 in the refolding of damaged proteins and in ER stress protection. Consistently, TRAP1 and/or TBP7 interference enhanced stress-induced cell death and increased intracellular protein ubiquitination. These experiments led us to hypothesize an involvement of TRAP1 in protein quality control for mistargeted/misfolded mitochondria-destined proteins, through interaction with the regulatory proteasome protein TBP7. Remarkably, expression of specific MITO proteins decreased upon TRAP1 interference as a consequence of increased ubiquitination. The proposed TRAP1 network has an impact in vivo, as it is conserved in human colorectal cancers, is controlled by ER-localized TRAP1 interacting with TBP7 and provides a novel model of the ER-mitochondria crosstalk. [ABSTRACT FROM AUTHOR]
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- 2012
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24. The polarized epithelial phenotype is dominant in hybrids between polarized and unpolarized rat thyroid cell lines
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Zurzolo, C., primary, Gentile, R., additional, Mascia, A., additional, Garbi, C., additional, Polistina, C., additional, Aloj, L., additional, Avvedimento, V.E., additional, and Nitsch, L., additional
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- 1991
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25. The tissue-specific pathways regulating cell proliferation are inherited independently in somatic hybrid between thyroid and liver cells.
- Author
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Veneziani, B M, primary, Villone, G, additional, Romano, R, additional, Di Carlo, A, additional, Garbi, C, additional, and Tramontano, D, additional
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- 1990
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26. The localization and activity of cAMP-dependent protein kinase affect cell cycle progression in thyroid cells.
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Feliciello, A, Gallo, A, Mele, E, Porcellini, A, Troncone, G, Garbi, C, Gottesman, M E, and Avvedimento, E V
- Abstract
cAMP signals are received and transmitted by multiple isoforms of cAMP-dependent protein kinases (PKAs), typically determined by their specific regulatory subunits. We describe changes in the cAMP signal transduction pathway during cell cycle progression in synchronized rat thyroid cells. Both PKA type II (PKAII) localization and nuclear cAMP signaling are significantly modified during G(0) and G(1)-S transitions. G(1) is characterized by PKA activation and amplified cAMP signal transduction. This is associated with a decrease in the concentration of RI and RII regulatory subunits and enhanced anchoring of PKAII to the Golgi-centrosome region. Just prior to S, the cAMP pathway is depressed. Up-regulation of the pathway by exogenous cAMP in G(1) inhibited the subsequent decay of the Cdk inhibitor p27 and delayed the onset of S phase. Forced translocation of endogenous PKAII to the cytosol down-regulated cAMP signaling, advancing the timing of p27 decay and inducing premature exit from G(1). These data indicate that membrane-bound PKA amplifies the transduction of cAMP signals in G(1) and that the length of G(1) is influenced by cAMP-PKA.
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- 2000
27. Change of inverted thyroid follicles into spheroid after embedding in a collagen gel
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GARBI C., TACCHETTI, CARLO, WOLLMAN S. H., Garbi, C., Tacchetti, Carlo, and Wollman, S. H.
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- 1986
28. Aspects of control of polarity of thyroid epithelial cells in follicles in suspension culture
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Wollman S. H., Nitsch L., Garbi C., TACCHETTI, CARLO, Eggo G.N., Burrow M.C., Wollman, S. H., Nitsch, L., Garbi, C., and Tacchetti, Carlo
- Published
- 1985
29. RhoA activity is required for fibronectin assembly and counteracts beta1B integrin inhibitory effect in FRT epithelial cells.
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Calì, G, Mazzarella, C, Chiacchio, M, Negri, R, Retta, S F, Zannini, M, Gentile, F, Tarone, G, Nitsch, L, and Garbi, C
- Abstract
FRT thyroid epithelial cells synthesize fibronectin and organize a network of fibronectin fibrils at the basal surface of the cells. Fibronectin fibril formation is enhanced by the overexpression of the ubiquitous beta1A integrin and is inhibited by the expression of the dominant-negative beta1B subunit. We tested the hypotheses that RhoA activity might mediate the integrin-dependent fibronectin fibrillogenesis and might counteract beta1B integrin inhibitory effect. FRT-beta1A cells were transfected with a vector carrying a dominant negative form of RhoA (RhoAN19) or treated with the C3 transferase exoenzyme. Both treatments inhibited fibronectin assembly and caused loss of actin microfilaments and adhesion plaques. On the other hand, FRT-beta1B cells were transfected with the constitutively activated form of RhoA (RhoAV14) or treated with the E. coli cytotoxic necrotizing factor 1, which directly activates RhoA. Either treatment restored microfilament and adhesion plaque assembly and promoted fibronectin fibril organization. A great increase in fibronectin fibril assembly was also obtained by treatment of FRT-beta1B cells with TGF-beta. Our data indicate that RhoA is required to promote fibronectin matrix assembly in FRT cells and that the activation of the signal transduction pathway downstream of RhoA can overcome the inhibitory effect of beta1B integrin.
- Published
- 1999
30. Yotiao protein, a ligand for the NMDA receptor, binds and targets cAMP-dependent protein kinase II^1
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Feliciello, A., Cardone, L., Garbi, C., Ginsberg, M.D., Varrone, S., Rubin, C.S., Avvedimento, E.V., and Gottesman, M.E.
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- 1999
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31. Basal lamina formation on thyroid epithelia in separated follicles in suspension culture.
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Garbi, C and Wollman, S H
- Abstract
When thyroid follicles are isolated by collagenase treatment of minced thyroid lobes, the basal lamina around each follicle is removed. The basal lamina does not reform when follicles are cultured in suspension in Coon's modified Ham's F-12 medium containing, in addition, 0.5% calf serum, insulin, transferrin, and thyrotropin. We have added acid soluble collagen and/or laminin to see if they would result in the formation of a basal lamina. An extended basal lamina did not form when follicles were embedded in a gel formed from acid-soluble rat tendon collagen or from calf skin collagen when added at a concentration of 100 micrograms collagen/ml. However, laminin at a concentration of 5.1 micrograms/ml gave rise to short segments of a basal lamina within 30 min. At longer time intervals, the segments lengthened and covered the base of many cells, and were continuous across the gap between cells and across the mouth of a coated pit. Not all basal surfaces were covered, and no exposed apical surfaces with microvilli had a basal lamina. There was no obvious difference in the appearance of the basal lamina if collagen was added in addition to laminin, but collagen, in contact with the plasma membrane when added alone, was lifted off the membrane in the presence of the basal lamina. The basal lamina appeared denser if formed in the presence of 5% serum instead of 0.5%.
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- 1982
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32. Cell fate following ER stress: Just a matter of 'quo ante' recovery or death?
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Treglia, A. S., Turco, S., Ulianich, L., Ausiello, P., dario domenico lofrumento, Nicolardi, G., Miele, C., Garbi, C., Beguinot, F., Di Jeso, B., A. S., Treglia, S., Turco, L., Ulianich, Ausiello, Pietro, D. D., Lofrumento, G., Nicolardi, C., Miele, Garbi, Corrado, Beguinot, Francesco, B. D., Jeso, Treglia, Antonella Sonia, Turco, Stefano, Ulianich, L, Ausiello, P, Lofrumento, Dario Domenico, Nicolardi, Giuseppe, Miele, C, Garbi, C, Beguinot, F, and DI JESO, Bruno
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PERK ,cell fate ,Apoptosis ,Recovery of Function ,Cell Dedifferentiation ,Endoplasmic Reticulum ,6 - Ciencias aplicadas::61 - Medicina [CDU] ,Stress, Physiological ,IRE ,Unfolded Protein Response ,Animals ,Humans ,ER stre ,Signal Transduction - Abstract
The endoplasmic reticulum (ER) is a complex and multifunctional organelle. It is the intracellular compartment of protein folding, a complex task, both facilitated and monitored by ER folding enzymes and molecular chaperones. The ER is also a stress-sensing organelle. It senses stress caused by disequilibrium between ER load and folding capacity and responds by activating signal transduction pathways, known as unfolded protein response (UPR). Three major classes of transducer are known, inositol-requiring protein-1 (IRE1), activating transcription factor-6 (ATF6), and protein kinase RNA (PKR)-like endoplasmic reticulum kinase (PERK), which sense with their endoluminal domain the state of protein folding, although the exact mechanism(s) involved is not entirely clear. Depending on whether the homeostatic response of the UPR is successful in restoring an equilibrium between ER load and protein folding or not, the two possible outcomes of the UPR so far considered have been life or death. Indeed, recent efforts have been devoted to understand the life/death switch mechanisms. However, recent data suggest that what appears to be a pure binary decision may in fact be more complex, and survival may be achieved at the expenses of luxury cell functions, such as expression of differentiation genes.
33. Increased hexosamine biosynthetic pathway flux alters cell-cell adhesion in INS-1E cells and murine islets
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Di Jeso, B., Turco, S., La Pesa, V., Treglia, A. S., dario domenico lofrumento, Nuccio, F., Ulianich, L., Miele, C., Longo, M., Spinelli, R., Parrillo, L., Nicolardi, G., Garbi, C., Beguinot, F., DI JESO, Bruno, S., Turco, V., La Pesa, Treglia, Antonella Sonia, Lofrumento, Dario Domenico, DE NUCCIO, Francesco, L., Ulianich, C., Miele, M., Longo, R., Spinelli, L., Parrillo, Nicolardi, Giuseppe, C., Garbi, and F., Beguinot
34. The TBC1D31/praja2 complex controls primary ciliogenesis through PKA‐directed OFD1 ubiquitylation
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Corrado Garbi, Bianca Fiorillo, Eduard Stefan, Brunella Franco, Manuela Morleo, Daniela Intartaglia, Omar Torres-Quesada, Emanuela Senatore, Ivan Conte, Bruno Catalanotti, Federica Moraca, Laura Rinaldi, Giuliana Giamundo, Marcel Kwiatkowski, Alienke van Pijkeren, Antonio Feliciello, Rossella Delle Donne, Andrea Raffeiner, Francesco Chiuso, Giovanni Scala, Luciano Pirone, Emilia Pedone, Senatore, E, Chiuso, F, Rinaldi, L, Intartaglia, D, Delle Donne, R, Pedone, E, Catalanotti, B, Pirone, L, Fiorillo, B, Moraca, F, Giamundo, G, Scala, G, Raffeiner, A, Torres-Quesada, O, Stefan, E, Kwiatkowski, M, van Pijkeren, A, Morleo, M, Franco, B, Garbi, C, Conte, I, Feliciello, A, Senatore, E., Chiuso, F., Rinaldi, L., Intartaglia, D., Delle Donne, R., Pedone, E., Catalanotti, B., Pirone, L., Fiorillo, B., Moraca, F., Giamundo, G., Scala, G., Raffeiner, A., Torres-Quesada, O., Stefan, E., Kwiatkowski, M., van Pijkeren, A., Morleo, M., Franco, B., Garbi, C., Conte, I., and Feliciello, A.
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Ubiquitin-Protein Ligases ,Oryzias ,Ciliopathies ,Article ,General Biochemistry, Genetics and Molecular Biology ,praja2 ,03 medical and health sciences ,0302 clinical medicine ,Ubiquitin ,Two-Hybrid System Techniques ,Ciliogenesis ,medicine ,Animals ,Humans ,PKA ,Membrane & Intracellular Transport ,Protein kinase A ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,General Immunology and Microbiology ,biology ,General Neuroscience ,Cilium ,Ubiquitination ,Post-translational Modifications, Proteolysis & Proteomics ,Articles ,medicine.disease ,Cyclic AMP-Dependent Protein Kinases ,Ubiquitin ligase ,Cell biology ,Ciliopathy ,Centrosome ,biology.protein ,OFD1 ,030217 neurology & neurosurgery ,primary cilium ,Signal Transduction - Abstract
The primary cilium is a microtubule‐based sensory organelle that dynamically links signalling pathways to cell differentiation, growth, and development. Genetic defects of primary cilia are responsible for genetic disorders known as ciliopathies. Orofacial digital type I syndrome (OFDI) is an X‐linked congenital ciliopathy caused by mutations in the OFD1 gene and characterized by malformations of the face, oral cavity, digits and, in the majority of cases, polycystic kidney disease. OFD1 plays a key role in cilium biogenesis. However, the impact of signalling pathways and the role of the ubiquitin‐proteasome system (UPS) in the control of OFD1 stability remain unknown. Here, we identify a novel complex assembled at centrosomes by TBC1D31, including the E3 ubiquitin ligase praja2, protein kinase A (PKA), and OFD1. We show that TBC1D31 is essential for ciliogenesis. Mechanistically, upon G‐protein‐coupled receptor (GPCR)‐cAMP stimulation, PKA phosphorylates OFD1 at ser735, thus promoting OFD1 proteolysis through the praja2‐UPS circuitry. This pathway is essential for ciliogenesis. In addition, a non‐phosphorylatable OFD1 mutant dramatically affects cilium morphology and dynamics. Consistent with a role of the TBC1D31/praja2/OFD1 axis in ciliogenesis, alteration of this molecular network impairs ciliogenesis in vivo in Medaka fish, resulting in developmental defects. Our findings reveal a multifunctional transduction unit at the centrosome that links GPCR signalling to ubiquitylation and proteolysis of the ciliopathy protein OFD1, with important implications on cilium biology and development. Derangement of this control mechanism may underpin human genetic disorders., OFD1 (oro‐facial digital type I syndrome protein) resides in a centrosomal complex that links GPCR signalling to ubiquitylation and degradation of OFD1, controlling cilium morphology and dynamics and vertebrate development.
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- 2021
35. Impact of Magnetic Stimulation on Periodontal Ligament Stem Cells
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Valentina Peluso, Laura Rinaldi, Teresa Russo, Olimpia Oliviero, Anna Di Vito, Corrado Garbi, Amerigo Giudice, Roberto De Santis, Antonio Gloria, Vincenzo D’Antò, Peluso, V., Rinaldi, L., Russo, T., Oliviero, O., Di Vito, A., Garbi, C., Giudice, A., De Santis, R., Gloria, A., and D'Anto, V.
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Adult ,magnetic stimulation design ,stem cells ,tissue engineering ,osteogenesis ,metabolomics ,cellular respiration ,Periodontal Ligament ,QH301-705.5 ,Cell Respiration ,Metabolomic ,Article ,Catalysis ,Inorganic Chemistry ,Young Adult ,Adenosine Triphosphate ,Humans ,Physical and Theoretical Chemistry ,Biology (General) ,Molecular Biology ,QD1-999 ,Spectroscopy ,Cell Proliferation ,Stem cell ,Osteogenesi ,Organic Chemistry ,Cell Differentiation ,General Medicine ,Alkaline Phosphatase ,equipment and supplies ,Mitochondria ,Computer Science Applications ,Chemistry ,Magnetic Fields ,human activities - Abstract
The aim of this study was to evaluate the effect of a time-dependent magnetic field on the biological performance of periodontal ligament stem cells (PDLSCs). A Western blot analysis and Alamar Blue assay were performed to investigate the proliferative capacity of magnetically stimulated PDLSCs (PDLSCs MAG) through the study of the MAPK cascade (p-ERK1/2). The observation of ALP levels allowed the evaluation of the effect of the magnetic field on osteogenic differentiation. Metabolomics data, such as oxygen consumption rate (OCR), extracellular acidification rate (ECAR) and ATP production provided an overview of the PDLSCs MAG metabolic state. Moreover, the mitochondrial state was investigated through confocal laser scanning microscopy. Results showed a good viability for PDLSCs MAG. Magnetic stimulation can activate the ERK phosphorylation more than the FGF factor alone by promoting a better cell proliferation. Osteogenic differentiation was more effectively induced by magnetic stimulation. The metabolic panel indicated significant changes in the mitochondrial cellular respiration of PDLSCs MAG. The results suggested that periodontal ligament stem cells (PDLSCs) can respond to biophysical stimuli such as a time-dependent magnetic field, which is able to induce changes in cell proliferation and differentiation. Moreover, the magnetic stimulation also produced an effect on the cell metabolic profile. Therefore, the current study demonstrated that a time-dependent magnetic stimulation may improve the regenerative properties of PDLSCs.
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- 2022
36. The Pervasive Effects of ER Stress on a Typical Endocrine Cell: Dedifferentiation, Mesenchymal Shift and Antioxidant Response in the Thyrocyte
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Gaetano Calì, Gregory Alexander Raciti, Corrado Garbi, Claudia Miele, Paola Mirra, Eduardo Consiglio, Antonella Sonia Treglia, Domenico Conza, Alessandro Miraglia, Francesco Beguinot, Bruno Di Jeso, Dario Punzi, Luca Ulianich, Ulianich, L., Mirra, P., Garbi, C., Cali, G., Conza, D., Treglia, A. S., Miraglia, A., Punzi, D., Miele, C., Raciti, G. A., Beguinot, F., Consiglio, E., and Di Jeso, B.
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0301 basic medicine ,Thyroid Epithelial Cell ,antioxidant response ,Endocrinology, Diabetes and Metabolism ,medicine.medical_treatment ,030209 endocrinology & metabolism ,Vimentin ,lcsh:Diseases of the endocrine glands. Clinical endocrinology ,Thyroglobulin ,Antioxidants ,thyroid ,Mesoderm ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,Downregulation and upregulation ,medicine ,Animals ,Endoplasmic Reticulum Stre ,Transcription factor ,Cells, Cultured ,Original Research ,lcsh:RC648-665 ,biology ,Chemistry ,Animal ,Endoplasmic reticulum ,Thyroid ,dedifferentiation ,mesenchymal phenotype ,Cell Differentiation ,Endoplasmic Reticulum Stress ,Cell biology ,Rats ,030104 developmental biology ,medicine.anatomical_structure ,Gene Expression Regulation ,Thyroid Epithelial Cells ,SNAI1 ,Unfolded protein response ,biology.protein ,Unfolded Protein Response ,ER stre ,Rat ,Antioxidant ,ER stress - Abstract
The endoplasmic reticulum stress and the unfolded protein response are triggered following an imbalance between protein load and protein folding. Until recently, two possible outcomes of the unfolded protein response have been considered: life or death. We sought to substantiate a third alternative, dedifferentiation, mesenchymal shift, and activation of the antioxidant response by using typical endocrine cells, i.e. thyroid cells. The thyroid is a unique system both of endoplasmic reticulum stress (a single protein, thyroglobulin represents the majority of proteins synthesized in the endoplasmic reticulum by the thyrocyte) and of polarized epithelium (the single layer of thyrocytes delimiting the follicle). Following endoplasmic reticulum stress, in thyroid cells the folding of thyroglobulin was disrupted. The mRNAs of unfolded protein response were induced or spliced (X-box binding protein-1). Differentiation was inhibited: mRNA levels of thyroid specific genes, and of thyroid transcription factors were dramatically downregulated, at least in part, transcriptionally. The dedifferentiating response was accompanied by an upregulation of mRNAs of antioxidant genes. Moreover, cadherin-1, and the thyroid (and kidney)-specific cadherin-16 mRNAs were downregulated, vimentin, and SNAI1 mRNAs were upregulated. In addition, loss of cortical actin and stress fibers formation were observed. Together, these data indicate that ER stress in thyroid cells induces dedifferentiation, loss of epithelial organization, shift towards a mesenchymal phenotype, and activation of the antioxidant response, highlighting, at the same time, a new and wide strategy to achieve survival following ER stress, and, as a sort of the other side of the coin, a possible new molecular mechanism of decline/loss of function leading to a deficit of thyroid hormones formation.
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- 2020
37. Feedback inhibition of cAMP effector signaling by a chaperone-assisted ubiquitin system
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Nicola Antonino Russo, Matthis Synofzik, Omar Torres-Quesada, Sonia Piccinin, Federica Moraca, Herbert Lindner, Bruno Catalanotti, Antonio Feliciello, Ulrich Stelzl, Verena Bachmann, Rossella Delle Donne, Laura Rinaldi, Corrado Garbi, Robert Nisticò, Antonella Scorziello, Lucio Annunziato, Francesco Chiuso, Eduard Stefan, Florian Enzler, Rinaldi, L., Delle Donne, R., Catalanotti, B., Torres-Quesada, O., Enzler, F., Moraca, F., Nistico', ROBERT GIOVANNI, Chiuso, Francesco, Piccinin, S., Bachmann, V., Lindner, H. H., Garbi, C., Scorziello, A., Russo, N. A., Synofzik, M., Stelzl, U., Annunziato, L., Stefan, Eduard, and Feliciello, A.
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0301 basic medicine ,pathology [Spinocerebellar Ataxias] ,Molecular Chaperone ,Ubiquitin-Protein Ligase ,Ubiquitylation ,Leupeptins ,metabolism [Cyclic AMP-Dependent Protein Kinase Catalytic Subunits] ,benzyloxycarbonylleucyl-leucyl-leucine aldehyde ,General Physics and Astronomy ,02 engineering and technology ,drug effects [Feedback, Physiological] ,Hippocampus ,Mice ,Ubiquitin ,HEK293 Cell ,Cyclic AMP ,genetics [Spinocerebellar Ataxias] ,antagonists & inhibitors [HSP70 Heat-Shock Proteins] ,Proteolysi ,Phosphorylation ,Receptor ,lcsh:Science ,metabolism [Molecular Chaperones] ,genetics [Ubiquitin-Protein Ligases] ,Feedback, Physiological ,HSP70 Heat-Shock Protein ,Multidisciplinary ,biology ,Chemistry ,Effector ,pharmacology [Purine Nucleosides] ,Settore BIO/14 ,021001 nanoscience & nanotechnology ,3. Good health ,Ubiquitin ligase ,Cell biology ,VER 155008 ,pharmacology [Leupeptins] ,Holoenzyme ,drug effects [Protein Binding] ,Fibroblast ,ddc:500 ,0210 nano-technology ,metabolism [Cyclic AMP] ,Stub1 protein, mouse ,Human ,Cell signalling ,Protein Binding ,Signal Transduction ,drug effects [Signal Transduction] ,congenital, hereditary, and neonatal diseases and abnormalities ,Science ,Ubiquitin-Protein Ligases ,Primary Cell Culture ,Leupeptin ,Purine Nucleoside ,General Biochemistry, Genetics and Molecular Biology ,metabolism [Holoenzymes] ,Article ,03 medical and health sciences ,metabolism [Ubiquitin-Protein Ligases] ,Hippocampu ,physiology [Signal Transduction] ,Animals ,Humans ,Spinocerebellar Ataxias ,HSP70 Heat-Shock Proteins ,Protein kinase A ,STUB1 protein, human ,Spinocerebellar Ataxia ,Cyclic AMP-Dependent Protein Kinase Catalytic Subunits ,Animal ,HEK 293 cells ,drug effects [Proteolysis] ,Ubiquitination ,General Chemistry ,Purine Nucleosides ,Fibroblasts ,physiology [Feedback, Physiological] ,Mice, Inbred C57BL ,030104 developmental biology ,pathology [Hippocampus] ,HEK293 Cells ,Ubiquitin ligases ,Chaperone (protein) ,Proteolysis ,biology.protein ,lcsh:Q ,Cyclic AMP-Dependent Protein Kinase Catalytic Subunit ,Holoenzymes ,physiology [Ubiquitination] ,Molecular Chaperones - Abstract
Activation of G-protein coupled receptors elevates cAMP levels promoting dissociation of protein kinase A (PKA) holoenzymes and release of catalytic subunits (PKAc). This results in PKAc-mediated phosphorylation of compartmentalized substrates that control central aspects of cell physiology. The mechanism of PKAc activation and signaling have been largely characterized. However, the modes of PKAc inactivation by regulated proteolysis were unknown. Here, we identify a regulatory mechanism that precisely tunes PKAc stability and downstream signaling. Following agonist stimulation, the recruitment of the chaperone-bound E3 ligase CHIP promotes ubiquitylation and proteolysis of PKAc, thus attenuating cAMP signaling. Genetic inactivation of CHIP or pharmacological inhibition of HSP70 enhances PKAc signaling and sustains hippocampal long-term potentiation. Interestingly, primary fibroblasts from autosomal recessive spinocerebellar ataxia 16 (SCAR16) patients carrying germline inactivating mutations of CHIP show a dramatic dysregulation of PKA signaling. This suggests the existence of a negative feedback mechanism for restricting hormonally controlled PKA activities., How intracellular cAMP activate PKA is well-characterized, but PKA inactivation remains poorly understood. Here, Rinaldi et al. show that CHIP/HSP70 ubiquitinates the catalytic subunit of PKA, with implications for the human disease spinocerebellar ataxia 16, as patients often have CHIP mutations.
- Published
- 2019
38. miR-34 modulates apoptotic gene expression in Ingenol mebutate treated keloid fibroblasts
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Margherita Santoriello, Corrado Garbi, Bruna De Felice, Massimo Nacca, Francesco Manfellotto, De Felice, B, Manfellotto, F, Garbi, C, Santoriello, M, and Nacca, M.
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p53 ,0301 basic medicine ,Cancer Research ,Cell ,DNA Fragmentation ,Biology ,Biochemistry ,03 medical and health sciences ,Keloid ,microRNA-34a ,microRNA ,Gene expression ,Genetics ,medicine ,Humans ,skin and connective tissue diseases ,Fibroblast ,Molecular Biology ,Cells, Cultured ,keloids ,apoptosis ,Apoptosi ,Articles ,Fibroblasts ,Cell cycle ,medicine.disease ,MicroRNAs ,030104 developmental biology ,medicine.anatomical_structure ,Gene Expression Regulation ,Oncology ,MicroRNA 34a ,ingenol mebutate ,Cancer research ,Molecular Medicine ,Apoptotic signaling pathway ,Diterpenes - Abstract
Keloids are benign skin tumors that develop in individuals who have a positive family history of keloid disorders. Keloids are characterized by a deregulated wound-healing process, atypical fibroblasts with extreme deposition of extracellular matrix components, particularly collagen, increased cell proliferation and associated failure of apoptosis. Recently ingenol-mebutate has been used as a novel agent with anti-proliferative activity on human keloids as an alternative treatment option in patients, once conventional therapies have failed. We hypothesized that microRNAs (miR/miRNA) may be involved in the balance between lesion formation and repair. A comprehensive understanding of the molecular mechanism underlying the Ingenol-mebutate response in keloid fibroblast following Ingenol-mebutate exposure has been established previously. Therefore, the present study analyzed changes in miRNAs and apoptotic gene regulation in Ingenol-mebutate treated keloid fibroblast, by reverse transcription-quantitative polymerase chain reaction and a DNA fragmentation assay. The range of upregulated miRNAs and downregulated genes encoding cell death appeared to be associated with the degree of the morphological alterations in Ingenol-mebutate treated keloids. In particular, the upregulation of miR-34a was detected in keloid fibroblasts during and following Ingenol-mebutate exposure. Keloid fibroblasts that overexpressed miR-34a showed differential expression of genes involved in the apoptotic signaling pathway such as p53. In conclusion, the Ingenol-mebutate treatment used here was effective in reducing keloid fibroblast growth in cell culture experiments and the expression of particular miRNAs modulated the pro-apoptotic gene expression following Ingenol-mebutate treatment.
- Published
- 2018
39. Increased hexosamine biosynthetic pathway flux dedifferentiates INS-1E cells and murine islets by an extracellular signal-regulated kinase (ERK)1/2-mediated signal transmission pathway
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Luca Ulianich, Giuseppe Nicolardi, Dario Domenico Lofrumento, Francesco Beguinot, Claudia Miele, Antonella Sonia Treglia, Corrado Garbi, Cecilia Nigro, B. Di Jeso, Luca Parrillo, Angela Lombardi, Lombardi, Angela, Ulianich, L, Treglia, Antonella Sonia, Nigro, C, Parrillo, L, Lofrumento, Dario Domenico, Nicolardi, Giuseppe, Garbi, C, Beguinot, F, Miele, C, DI JESO, Bruno, Lombardi, A, Ulianich, Luca, Treglia, A. S, Nigro, Cecilia, Parrillo, Luca, Lofrumento, D. D, Nicolardi, Federica, Garbi, Corrado, Beguinot, Francesco, and Miele, Claudia
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Beta cells Dedifferentiation ERK1/2 ER stress ,MAPK/ERK pathway ,MAP Kinase Signaling System ,Endocrinology, Diabetes and Metabolism ,Protein Kinase Inhibitor ,Down-Regulation ,Biology ,medicine.disease_cause ,Cell Line ,Clone Cell ,Islets of Langerhans ,Mice ,Downregulation and upregulation ,Insulin Secretion ,Internal Medicine ,medicine ,Animals ,Insulin ,RNA, Messenger ,Phosphorylation ,Endoplasmic Reticulum Stre ,Phenylbutyrate ,Protein Kinase Inhibitors ,Homeodomain Proteins ,Mitogen-Activated Protein Kinase 1 ,Glucosamine ,Mitogen-Activated Protein Kinase 3 ,Animal ,Endoplasmic reticulum ,Homeodomain Protein ,Islets of Langerhan ,Cell Dedifferentiation ,Endoplasmic Reticulum Stress ,Phenylbutyrates ,Clone Cells ,Rats ,Cell biology ,Mice, Inbred C57BL ,Trans-Activator ,Apoptosis ,Trans-Activators ,Unfolded Protein Response ,Unfolded protein response ,Rat ,Beta cell ,Protein Processing, Post-Translational ,Flux (metabolism) ,Oxidative stress - Abstract
AIMS/HYPOTHESIS: Beta cell failure is caused by loss of cell mass, mostly by apoptosis, but also by simple dysfunction (decline of glucose-stimulated insulin secretion, downregulation of specific gene expression). Apoptosis and dysfunction are caused, at least in part, by lipoglucotoxicity. The mechanisms implicated are oxidative stress, increase in the hexosamine biosynthetic pathway (HBP) flux and endoplasmic reticulum (ER) stress. Oxidative stress plays a role in glucotoxicity-induced beta cell dedifferentiation, while glucotoxicity-induced ER stress has been mostly linked to beta cell apoptosis. We sought to clarify whether ER stress caused by increased HBP flux participates in a dedifferentiating response of beta cells, in the absence of relevant apoptosis. METHODS: We used INS-1E cells and murine islets. We analysed the unfolded protein response and the expression profile of beta cells by real-time RT-PCR and western blot. The signal transmission pathway elicited by ER stress was investigated by real-time RT-PCR and immunofluorescence. RESULTS: Glucosamine and high glucose induced ER stress, but did not decrease cell viability in INS-1E cells. ER stress caused dedifferentiation of beta cells, as shown by downregulation of beta cell markers and of the transcription factor, pancreatic and duodenal homeobox 1. Glucose-stimulated insulin secretion was inhibited. These effects were prevented by the chemical chaperone, 4-phenyl butyric acid. The extracellular signal-regulated kinase (ERK) signal transmission pathway was implicated, since its inhibition prevented the effects induced by glucosamine and high glucose. CONCLUSIONS/INTERPRETATION: Glucotoxic ER stress dedifferentiates beta cells, in the absence of apoptosis, through a transcriptional response. These effects are mediated by the activation of ERK1/2.
- Published
- 2011
40. Effect of selenocystine on gene expression profiles in human keloid fibroblasts
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Margherita Santoriello, Massimo Nacca, Bruna De Felice, Corrado Garbi, Robert R. Wilson, DE FELICE, Bruna, Garbi, C, Wilson, Rr, Santoriello, M, Nacca, M., De Felice, B., Garbi, Corrado, Wilson, R. R., and Santoriello, Margherita
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Keloids, * Microarray, * Selenocystine, * Gene expression ,Programmed cell death ,Microarray ,Selenocystine ,Biology ,Extracellular matrix ,Keloid ,Gene expression ,medicine ,Genetics ,Humans ,Fibroblast ,Cell adhesion ,skin and connective tissue diseases ,Transcription factor ,Cells, Cultured ,Oligonucleotide Array Sequence Analysis ,Reverse Transcriptase Polymerase Chain Reaction ,Gene Expression Profiling ,Proteins ,Fibroblasts ,medicine.disease ,Selenocysteine ,medicine.anatomical_structure ,Gene Expression Regulation ,Immunology ,Keloids ,Cancer research - Abstract
In this study, selenocystine, a nutritionally available selenoamino acid, was identified for the first time as a novel agent with anti proliferative activity on human keloids. The 20 μM concentration after 48. h treatment used here was the most effective to reduce keloid fibroblast growth. We analyzed the gene expression profile of selenocystine treatment response in keloid fibroblasts by the microarray system to characterize the effects of selenocystine on human keloids. The major alterations in keloid fibroblasts following selenocystine exposure included up-regulation of the genes encoding cell death and transcription factors. Prominent down-regulation of genes involved in development, cell adhesion and cytoskeleton, as well as extra cellular matrix genes, usually strongly up-regulated in keloids, resulted following selenocystine exposure. The range of the down-regulated genes and the degree of the decreased expression appeared to be correlated with the degree of the morphological alterations in selenocystine treated keloids. © 2011 Elsevier Inc.
- Published
- 2011
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41. Tyr Phosphatase-Mediated P-ERK Inhibition Suppresses Senescence in EIA + v-raf Transformed Cells, Which, Paradoxically, Are Apoptosis-Protected in a MEK-Dependent Manner
- Author
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Angela Lombardi, Stefania De Vitis, Corrado Garbi, Francesco Beguinot, Claudia Miele, Giuseppe Terrazzano, Antonella Sonia Treglia, Luca Ulianich, Stefano Turco, Bruno Di Jeso, De Vitis, S., Treglia, S. A., Ulianich, L., Turco, S., Terrazzano, G., Lombardi, A., Miele, C., Garbi, Corrado, Beguinot, Francesco, Di Jeso, B., De Vitis, S, Treglia, Antonella Sonia, Ulianich, L, Turco, Stefano, Terrazzano, G, Lombardi, A, Miele, C, Garbi, C, Beguinot, F, and DI JESO, Bruno
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MAPK/ERK pathway ,Cancer Research ,Phosphatase ,MELANOMA ,Protein tyrosine phosphatase ,Biology ,Mitogen-activated protein kinase kinase ,lcsh:RC254-282 ,ACTIVATION ,Annexin ,A-RAF ,thyroid cancer ,phosphatases ,Protein kinase A ,Kinase ,MUTATIONS ,MAP KINASE ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,Molecular biology ,CANCER ,PROTEIN-TYROSINE PHOSPHATASES ,C-RAF ,ERK ,KINASE-ACTIVITY ,GROWTH ,Cell aging - Abstract
Activation of the Ras-Raf–extracellular signal–regulated kinase (ERK) pathway causes not only proliferation and suppression of apoptosis but also the antioncogenic response of senescence. How these contrasting effects are reconciled to achieve cell transformation and cancer formation by this pathway is poorly understood. In a systemof two-step carcinogenesis (dedifferentiated PC EIA, transformed PC EIA–polyoma–middle T [PC EIA + Py] and PC EIA–v-raf [PC EIA + raf] cells], v-raf cooperated with EIA by virtue of a strong prosurvival effect, not elicited by Py–middle T, evident toward serum-deprivation– and H2O2-induced apoptosis. Apoptosis was detected by DNA fragmentation and annexin V staining. The prosurvival function of v-raf was, in part, mitogen-activated protein kinase/ERK kinase (MEK)– dependent, as shown by pharmacologicalMEK inhibition. TheMEK-dependent antiapoptotic effect of v-raf was exerted despite a lower level of P-ERK1/2 in EIA + raf cells with respect to EIA + Py/EIA cells, which was dependent on a high tyrosine phosphatase activity, as shown by orthovanadate blockade. An ERK1/2 tyrosine phosphatase was likely involved. The high tyrosine phosphatase activity was instrumental to the complete suppression of senescence, detected by senescence-associated β-galactosidase activity, because tyrosine phosphatase blockade induced senescence in EIA + raf but not in EIA + Py cells. High tyrosine phosphatase activity and evasion from senescence were confirmed in an anaplastic thyroid cancer cell line. Therefore, besides EIA, EIA + raf cells suppress senescence through a new mechanism, namely, phosphatase-mediated P-ERK1/2 inhibition, but, paradoxically, retain the oncogenic effects of the Raf-ERK pathway. We propose that the survival effect of Raf is not a function of absolute P-ERK1/2 levels at a given time but is rather dynamically dependent on greater variations after an apoptotic stimulus.
- Published
- 2011
42. T cell activation induces CuZn superoxide dismutase (SOD)-1 intracellular re-localization, production and secretion
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Anna Sasso, Valentina Ucci, Giuseppina Ruggiero, Bruna De Felice, Angela Giovazzino, Mariarosaria Santillo, Simona Damiano, Paolo Mondola, Tiziana Petrozziello, Anna Teresa Palatucci, Corrado Garbi, Valentina Rubino, Giuseppe Terrazzano, Terrazzano, G, Rubino, Valentina, Damiano, S, Sasso, A, Petrozziello, T, Ucci, V, Palatucci, At, Giovazzino, A, Santillo, Mariarosaria, De Felice, B, Garbi, Corrado, Mondola, Paolo, Ruggiero, Giuseppina, Rubino, V, Santillo, M, DE FELICE, Bruna, Garbi, C, Mondola, P, and Ruggiero, G.
- Subjects
Microvesicle secretion ,CD3 Complex ,Human T lymphocyte ,T cell ,T-Lymphocytes ,SOD-1 ,Intracellular Space ,Receptors, Antigen, T-Cell ,Activation ,Lymphocyte Activation ,Superoxide dismutase ,chemistry.chemical_compound ,Superoxide Dismutase-1 ,TCR triggering ,Extracellular ,medicine ,T lymphocyte ,Cytotoxic T cell ,Cluster Analysis ,Humans ,Molecular Biology ,Cell Aggregation ,chemistry.chemical_classification ,Reactive oxygen species ,Brefeldin A ,biology ,Superoxide dismutase (SOD1) ,Superoxide ,T cell activation ,Superoxide Dismutase ,T-cell receptor ,Cytoplasmic Vesicles ,ROS ,Cell Biology ,Cell biology ,Acetylcysteine ,Protein Transport ,medicine.anatomical_structure ,chemistry ,Intracellular localization ,Enzyme Induction ,biology.protein ,Reactive Oxygen Species ,Intracellular - Abstract
Reactive oxygen species (ROS) behave as second messengers in signal transduction for a series of receptor/ligand interactions. A major regulatory role is played by hydrogen peroxide (H2O2), more stable and able to freely diffuse through cell membranes. Copper-zinc superoxide dismutase (CuZn-SOD)-1 is a cytosolic enzyme involved in scavenging oxygen radicals to H2O2 and molecular oxygen, thus representing a major cytosolic source of peroxides. Previous studies suggested that superoxide anion and H2O2 generation are involved in T cell receptor (TCR)-dependent signaling. Here, we describe that antigen-dependent activation of human T lymphocytes significantly increased extracellular SOD-1 levels in lymphocyte cultures. This effect was accompanied by the synthesis of SOD-1-specific mRNA and by the induction of microvesicle SOD-1 secretion. It is of note that SOD-1 increased its concentration specifically in T cell population, while no significant changes were observed in the "non-T" cell counterpart. Moreover, confocal microscopy showed that antigen-dependent activation was able to modify SOD-1 intracellular localization in T cells. Indeed, was observed a clear SOD-1 recruitment by TCR clusters. The ROS scavenger N-acetylcysteine (NAC) inhibited this phenomenon. Further studies are needed to define whether SOD-1-dependent superoxide/peroxide balance is relevant for regulation of T cell activation, as well as in the functional cross talk between immune effectors. © 2013 Elsevier B.V.
- Published
- 2013
43. ER stress is associated with dedifferentiation and an epithelial-to-mesenchymal transition-like phenotype in PC Cl3 thyroid cells
- Author
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Francesco Beguinot, Claudia Miele, Gregory Alexander Raciti, Bruno Di Jeso, Antonella Sonia Treglia, Corrado Garbi, Dario Punzi, Eduardo Consiglio, Luca Ulianich, Ulianich, L, Garbi, C, Treglia, Antonella Sonia, Punzi, D, Miele, C, Raciti, Ga, Beguinot, F, Consiglio, E, DI JESO, Bruno, Ulianich, L., Garbi, Corrado, Treglia, A. S., Punzi, D., Miele, C., Raciti, G. A., Beguinot, Francesco, Consiglio, E., and Di Jeso, B.
- Subjects
tiroide ,TUMOR-CELLS ,medicine.medical_treatment ,Thyroid Gland ,Fluorescent Antibody Technique ,Vimentin ,Endoplasmic Reticulum ,Mesoderm ,transizione epitelio-mesenchimale ,chemistry.chemical_compound ,ENDOPLASMIC-RETICULUM STRESS ,TRANSCRIPTION FACTOR SNAIL ,biology ,UNFOLDED PROTEIN RESPONSE ,Tunicamycin ,Cell Differentiation ,dedifferenziazione ,Cadherins ,Cell biology ,DIFFERENTIATION ,Thapsigargin ,EPIDERMAL-GROWTH-FACTOR ,medicine.medical_specialty ,endocrine system ,Blotting, Western ,stress del reticolo endoplasmico ,Cell Line ,THYROGLOBULIN ,Thyroid peroxidase ,Internal medicine ,E-CADHERIN ,medicine ,Animals ,BREAST-CANCER ,RNA, Messenger ,MESSENGER-RNAS ,Endoplasmic reticulum ,Epithelial Cells ,Cell Biology ,Blotting, Northern ,Rats ,Endocrinology ,Gene Expression Regulation ,chemistry ,SNAI1 ,biology.protein ,Unfolded protein response ,Thyroglobulin - Abstract
Conditions perturbing the homeostasis of the endoplasmic reticulum (ER) cause accumulation of unfolded proteins and trigger ER stress. In PC Cl3 thyroid cells, thapsigargin and tunicamycin interfered with the folding of thyroglobulin, causing accumulation of this very large secretory glycoprotein in the ER. Consequently, mRNAs encoding BiP and XBP-1 were induced and spliced, respectively. In the absence of apoptosis, differentiation of PC Cl3 cells was inhibited. mRNA and protein levels of the thyroid-specific genes encoding thyroglobulin, thyroperoxidase and the sodium/iodide symporter and of the genes encoding the thyroid transcription factors TTF-1, TTF-2 and Pax-8 were dramatically downregulated. These effects were, at least in part, transcriptional. Moreover, they were selective and temporally distinct from the general and transient PERK-dependent translational inhibition. Thyroid dedifferentiation was accompanied by changes in the organization of the polarized epithelial monolayer. Downregulation of the mRNA encoding E-cadherin, and upregulation of the mRNAs encoding vimentin, α-smooth muscle actin, α(1)(I) collagen and SNAI1/SIP1, together with formation of actin stress fibers and loss of trans-epithelial resistance were found, confirming an epithelial-mesenchymal transition (EMT). The thyroid-specific and epithelial dedifferentiation by thapsigargin or tunicamycin were completely prevented by the PP2 inhibitor of Src-family kinases and by stable expression of a dominant-negative Src. Together, these data indicate that ER stress induces dedifferentiation and an EMT-like phenotype in thyroid cells through a Src-mediated signaling pathway.
- Published
- 2008
44. Analysis of codherin/catenin complexes in transformed thyroid epithelial cells: Modulation by beta 1 integrin subunit
- Author
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Corrado Garbi, Dario Greco, Lucio Nitsch, M. Grieco, Angela Celetti, Aniello Cerrato, Evelina Mele, Claudia Consales, Celetti, A, Garbi, C, Consales, C, Cerrato, A, Greco, D, Mele, E, Nitsch, L, Grieco, Michele, Garbi, Corrado, Nitsch, Lucio, and Grieco, M.
- Subjects
Histology ,Beta-catenin ,integrin ,Beta I integrin ,Blotting, Western ,Retroviridae Proteins, Oncogenic ,Genes, myc ,Thyroid Gland ,Alpha catenin ,Fluorescent Antibody Technique ,Gene Expression ,Cell Communication ,Oncogene Proteins v-raf ,Pathology and Forensic Medicine ,thyroid ,Sarcoma Viruses, Murine ,Cell Movement ,Animals ,beta Catenin ,Cell Line, Transformed ,Thyroid Epithelial Cells ,biology ,Cadherin ,Integrin beta1 ,Epithelial Cells ,Cell Biology ,General Medicine ,Cadherins ,Molecular biology ,catenins ,Rats ,Cell biology ,Catenin ,Cytoskeletal Proteins ,Genes, ras ,cadherin ,Desmoplakins ,Integrin alpha M ,Trans-Activators ,biology.protein ,beta 1 integrin ,Integrin, beta 6 ,Adenovirus E1A Proteins ,Collagen ,gamma Catenin ,Catenin complex ,Gels ,alpha Catenin - Abstract
We have analysed the expression of cadherin/catenin complex molecules in PC C13 rat thyroid cells transformed in vitro with different oncogenes, No significant downregulation of either E-cadherin, alpha-, beta- and gamma-catenin was detected following the introduction of activated forms of myc, adenovirus E1A, ras, raf, myc + ras, EIA + raf, However, ras- and raf-transformed PC C13 cells showed altered adherens junctions. An altered distribution of cadherin/catenin complexes characterized by radially oriented membrane spikes perpendicular to cell edges was the most prominent feature evidenced by immunofluorescence. No beta 1 integrin localization was observed in areas where this altered pattern of E-cadherin expression was detected. However, beta 1 integrin submit expression was detected at areas of cell-cell contact where E-cadherin showed a normal pattern of expression. Furthermore, ras- and raf-transformed PC C13 cells showed the ability to migrate in collagen gels, in contrast to their normal untransformed counterpart, Overexpression of beta 1 integrin was found to restore normal E-cadherin localization at cell-cell contacts and to partially inhibit the ability to migrate in collagen gels. Finally, two cell lines obtained by ras transformation in vivo, and derived from a rat primary thyroid carcinoma (TK6) and its lung metastasis (MPTK6), were found to have lost gamma-catenin expression. TK6 lost also E-cadherin expression and membrane localization of alpha-catenin, These results suggest that: i) in vitro thyroid cell transformation is associated to a change in cadherin/catenin complexes distribution rather than to a decrease in expression; ii) in vivo transformation is associated to the loss of expression of some of these molecules likely due to tumor progression; iii) alterations in beta 1 integrin subunit expression can result in changes in cadherin/catenin function thus implying that an integrin-cadherin synergy may exist in thyroid cells.
- Published
- 2000
45. Flow-cytometric detection of the RI alpha subunit of type I cAMP-dependent protein kinase in human cells
- Author
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Angelo Raffaele Bianco, Corrado Garbi, A. Ruggiero, Hiroshi Yokozaki, Giampaolo Tortora, Bjorn S. Skalhegg, Timothy Clair, Stefano Pepe, Fortunato Ciardiello, Yoon S. Cho-Chung, Pepe, S, Ruggiero, A, Tortora, G, Ciardiello, F, Garbi, Corrado, Yokozaki, H, CHO CHUNG, Y, Clair, T, Skalhegg, B, Bianco, Ar, Ciardiello, Fortunato, and Garbi, C
- Subjects
Protein subunit ,Cyclic AMP-Dependent Protein Kinase RIalpha Subunit ,Recombinant Fusion Proteins ,Immunofluorescence ,Biophysics ,Cyclic AMP-Dependent Protein Kinase Type II ,Biology ,Transfection ,Pathology and Forensic Medicine ,Flow cytometry ,Endocrinology ,human mammary cells ,medicine ,Humans ,Neoplastic transformation ,Breast ,Lymphocytes ,Protein kinase A ,Immunofluorescence, cAMPdependent protein kinase, cell cycle, human lymphocytes, human mammary cells ,Cells, Cultured ,G alpha subunit ,medicine.diagnostic_test ,Photoaffinity labeling ,Cell growth ,cAMPdependent protein kinase ,Cell Biology ,Hematology ,DNA ,Flow Cytometry ,Molecular biology ,Cyclic AMP-Dependent Protein Kinases ,human lymphocytes ,Biochemistry ,Cell culture ,Enzyme Induction ,cell cycle - Abstract
cAMP-dependent protein kinase (PKA) is composed of two genetically distinct catalytic (C) and regulatory (R) subunits. There are two different classes of PKA, designated as type I and type II, which contain distinct R subunits (RI or RII, respectively) but share a common C subunit. Enhanced expression of type I PKA has been correlated with cell proliferation and neoplastic transformation. Detection of the different PKA subunits is usually performed by photoaffinity labeling with 8-N3-32P-cAMP or by radioimmunolabeling techniques. Both techniques are time consuming and require a high number of cells and the use of radioactive reagents. Using the MCF-10A normal human mammary cell line infected with a recombinant retroviral vector containing the human RI alpha gene (MCF-10A RI alpha), we have developed a flow-cytometric assay to detect the intracellular content of RI alpha protein in human cells. MCF-10A and MCF-10A RI alpha cells were fixed in 1.5% paraformaldehyde at 37 degrees C for 15 min and permeabilized by methanol and acetone (1:1) at -20 degrees C for 5 min before staining with a specific IgG2a MoAb followed by a FITC-conjugate rabbit-anti mouse IgG. This procedure was also successfully utilized to recognize RI alpha protein content in human peripheral blood lymphocytes. Flow-cytometric detection of the RI alpha subunit in human cells is feasible and allows the study of the role of type I PKA in cell growth and neoplastic transformation.
- Published
- 1994
46. Author Correction: Feedback inhibition of cAMP effector signaling by a chaperone-assisted ubiquitin system.
- Author
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Rinaldi L, Donne RD, Catalanotti B, Torres-Quesada O, Enzler F, Moraca F, Nisticò R, Chiuso F, Piccinin S, Bachmann V, Lindner HH, Garbi C, Scorziello A, Russo NA, Synofzik M, Stelzl U, Annunziato L, Stefan E, and Feliciello A
- Published
- 2024
- Full Text
- View/download PDF
47. Downregulation of praja2 restrains endocytosis and boosts tyrosine kinase receptors in kidney cancer.
- Author
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Rinaldi L, Chiuso F, Senatore E, Borzacchiello D, Lignitto L, Iannucci R, Donne RD, Fuggi M, Reale C, Russo F, Russo NA, Giurato G, Rizzo F, Sellitto A, Santangelo M, De Biase D, Paciello O, D'Ambrosio C, Amente S, Garbi C, Dalla E, Scaloni A, Weisz A, Ambrosino C, Insabato L, and Feliciello A
- Subjects
- Adult, Animals, Humans, Mice, Down-Regulation, Endocytosis, ErbB Receptors genetics, ErbB Receptors metabolism, Proteasome Endopeptidase Complex metabolism, Receptor Protein-Tyrosine Kinases genetics, Ubiquitin metabolism, Carcinoma, Renal Cell genetics, Carcinoma, Renal Cell pathology, Kidney Neoplasms genetics, Kidney Neoplasms pathology
- Abstract
Clear cell renal cell carcinoma (ccRCC) is the most common kidney cancer in the adult population. Late diagnosis, resistance to therapeutics and recurrence of metastatic lesions account for the highest mortality rate among kidney cancer patients. Identifying novel biomarkers for early cancer detection and elucidating the mechanisms underlying ccRCC will provide clues to treat this aggressive malignant tumor. Here, we report that the ubiquitin ligase praja2 forms a complex with-and ubiquitylates the AP2 adapter complex, contributing to receptor endocytosis and clearance. In human RCC tissues and cells, downregulation of praja2 by oncogenic miRNAs (oncomiRs) and the proteasome markedly impairs endocytosis and clearance of the epidermal growth factor receptor (EGFR), and amplifies downstream mitogenic and proliferative signaling. Restoring praja2 levels in RCC cells downregulates EGFR, rewires cancer cell metabolism and ultimately inhibits tumor cell growth and metastasis. Accordingly, genetic ablation of praja2 in mice upregulates RTKs (i.e. EGFR and VEGFR) and induces epithelial and vascular alterations in the kidney tissue.In summary, our findings identify a regulatory loop between oncomiRs and the ubiquitin proteasome system that finely controls RTKs endocytosis and clearance, positively impacting mitogenic signaling and kidney cancer growth., (© 2024. The Author(s).)
- Published
- 2024
- Full Text
- View/download PDF
48. Carbonic anhydrase IX subcellular localization in normoxic and hypoxic SH-SY5Y neuroblastoma cells is assisted by its C-terminal protein interaction domain.
- Author
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Succoio M, Amiranda S, Sasso E, Marciano C, Finizio A, De Simone G, Garbi C, and Zambrano N
- Abstract
The human carbonic anhydrase IX (CA IX) is a hypoxia-induced transmembrane protein belonging to the α-CA enzyme family. It has a crucial role in pH regulation in hypoxic cells and acts by buffering intracellular acidosis induced by hypoxia. Indeed, it is frequently expressed in cancer cells, where it contributes to tumor progression. CA IX is also able to localize in the nucleus, where it contributes to 47S rRNA precursor genes transcription; however, the mechanisms assisting its nuclear translocation still remain unclear. The aim of our study was to deepen the understanding of the mechanisms involved in CA IX subcellular distribution. To this purpose, we implemented a site-directed mutagenesis approach targeting the C-terminal domain of CA IX and evaluated the subcellular distribution of the wild-type and mutant proteins in the SH-SY5Y cell line. The mutant proteins showed impaired binding ability and altered subcellular distribution in both normoxic and hypoxic conditions. Our data suggest that CA IX nuclear translocation depends on its transit through the secretory and the endocytic pathways., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)
- Published
- 2023
- Full Text
- View/download PDF
49. Ubiquitylation of BBSome is required for ciliary assembly and signaling.
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Chiuso F, Delle Donne R, Giamundo G, Rinaldi L, Borzacchiello D, Moraca F, Intartaglia D, Iannucci R, Senatore E, Lignitto L, Garbi C, Conflitti P, Catalanotti B, Conte I, and Feliciello A
- Subjects
- Animals, Protein Transport, Signal Transduction, Receptors, G-Protein-Coupled genetics, Ubiquitination, Cilia metabolism, Bardet-Biedl Syndrome genetics
- Abstract
Bardet-Biedl syndrome (BBS) is a ciliopathy characterized by retinal degeneration, obesity, renal abnormalities, postaxial polydactyly, and developmental defects. Genes mutated in BBS encode for components and regulators of the BBSome, an octameric complex that controls the trafficking of cargos and receptors within the primary cilium. Although both structure and function of the BBSome have been extensively studied, the impact of ubiquitin signaling on BBSome is largely unknown. We identify the E3 ubiquitin ligase PJA2 as a novel resident of the ciliary compartment and regulator of the BBSome. Upon GPCR-cAMP stimulation, PJA2 ubiquitylates BBSome subunits. We demonstrate that ubiquitylation of BBS1 at lysine 143 increases the stability of the BBSome and promotes its binding to BBS3, an Arf-like GTPase protein controlling the targeting of the BBSome to the ciliary membrane. Downregulation of PJA2 or expression of a ubiquitylation-defective BBS1 mutant (BBS1
K143R ) affects the trafficking of G-protein-coupled receptors (GPCRs) and Shh-dependent gene transcription. Expression of BBS1K143R in vivo impairs cilium formation, embryonic development, and photoreceptors' morphogenesis, thus recapitulating the BBS phenotype in the medaka fish model., (© 2023 The Authors. Published under the terms of the CC BY 4.0 license.)- Published
- 2023
- Full Text
- View/download PDF
50. Targeted inhibition of ubiquitin signaling reverses metabolic reprogramming and suppresses glioblastoma growth.
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Delle Donne R, Iannucci R, Rinaldi L, Roberto L, Oliva MA, Senatore E, Borzacchiello D, Lignitto L, Giurato G, Rizzo F, Sellitto A, Chiuso F, Castaldo S, Scala G, Campani V, Nele V, De Rosa G, D'Ambrosio C, Garbi C, Scaloni A, Weisz A, Ambrosino C, Arcella A, and Feliciello A
- Subjects
- Adult, Animals, Cell Line, Tumor, Humans, Mice, Proteasome Endopeptidase Complex metabolism, Signal Transduction, Ubiquitin, Brain Neoplasms genetics, Brain Neoplasms pathology, Glioblastoma metabolism
- Abstract
Glioblastoma multiforme (GBM) is the most frequent and aggressive form of primary brain tumor in the adult population; its high recurrence rate and resistance to current therapeutics urgently demand a better therapy. Regulation of protein stability by the ubiquitin proteasome system (UPS) represents an important control mechanism of cell growth. UPS deregulation is mechanistically linked to the development and progression of a variety of human cancers, including GBM. Thus, the UPS represents a potentially valuable target for GBM treatment. Using an integrated approach that includes proteomics, transcriptomics and metabolic profiling, we identify praja2, a RING E3 ubiquitin ligase, as the key component of a signaling network that regulates GBM cell growth and metabolism. Praja2 is preferentially expressed in primary GBM lesions expressing the wild-type isocitrate dehydrogenase 1 gene (IDH1). Mechanistically, we found that praja2 ubiquitylates and degrades the kinase suppressor of Ras 2 (KSR2). As a consequence, praja2 restrains the activity of downstream AMP-dependent protein kinase in GBM cells and attenuates the oxidative metabolism. Delivery in the brain of siRNA targeting praja2 by transferrin-targeted self-assembling nanoparticles (SANPs) prevented KSR2 degradation and inhibited GBM growth, reducing the size of the tumor and prolonging the survival rate of treated mice. These data identify praja2 as an essential regulator of cancer cell metabolism, and as a potential therapeutic target to suppress GBM growth., (© 2022. The Author(s).)
- Published
- 2022
- Full Text
- View/download PDF
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