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5. Additional file 1 of Expression of fatty acid synthase genes and their role in development and arboviral infection of Aedes aegypti

Author

Chotiwan, Nunya, Brito-Sierra, Carlos A., Ramirez, Gabriella, Lian, Elena, Grabowski, Jeffrey M., Graham, Babara, Hill, Catherine A., and Perera, Rushika

Abstract

Additional file 1: Figure S1. Alignment of the conserved YKELRLRGY motif. Figure S2. Amino acid alignment of the pseudo-methyltransferase (ΨME) domain of H. sapiens and Ae. aegypti FAS. Figure S3. RT-PCR assays designed to detect mRNA products of AaFAS-like and AaFAS6. Table S1. List of vertebrate, invertebrate and yeast FAS gene models employed in the present study. Table S2. Primers for generation of dsRNA for knock-down studies. Table S3. Primers for AaFAS expression analyses. Table S4. Primers for RT-PCR assay detecting mRNA products AaFAS-like and AaFAS6. Table S5. Amino acid similarity of FAS domains between H. sapiens and Ae. aegypti. Table S6. Comparisons of percent infection in mosquitoes (P-values). Table S7. Virus titers in mosquito tissues.

Published
2022
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11. BiP: Master Regulator of the Unfolded Protein Response and Crucial Factor in Flavivirus Biology

Author

Lewy, Tyler G., Grabowski, Jeffrey M., and Bloom, Marshall E.

Subjects
GRP78, Kunjin, BiP, tick-borne encephalitis virus, viruses, Flavivirus, fungi, macromolecular substances, Review, unfolded protein response, biochemical phenomena, metabolism, and nutrition, Virus Replication, complex mixtures, dengue, West Nile, Flavivirus Infections, Animals, Humans, Japanese encephalitis, Endoplasmic Reticulum Chaperone BiP, Heat-Shock Proteins
Abstract

Flaviviruses have an intimate relationship with their host cells, utilizing host proteins during replication. Much of viral genome replication and virion assembly occurs on and within the endoplasmic reticulum (ER). As a cellular protein folding hub, the ER provides an ideal environment for flaviviruses to replicate. Flaviviruses can interact with several ER processes, including the unfolded protein response (UPR), a cellular stress mechanism responsible for managing unfolded protein accumulation and ER stress. The UPR can alter the ER environment in several ways, including increasing ER volume and quantity of available chaperones, both of which can favor viral replication. BiP, a chaperone and master regulator of the UPR, has been demonstrated to play a key role in several flavivirus infections. Here we describe what is known in regard to BiP, its implicated role with flavivirus infection, and what remains to be discovered.

Published
2017

14. Dynamic remodeling of lipids coincides with dengue virus replication in the midgut of Aedes aegypti mosquitoes

Author

Chotiwan, Nunya, primary, Andre, Barbara G., additional, Sanchez-Vargas, Irma, additional, Islam, M. Nurul, additional, Grabowski, Jeffrey M., additional, Hopf-Jannasch, Amber, additional, Gough, Erik, additional, Nakayasu, Ernesto, additional, Blair, Carol D., additional, Belisle, John T., additional, Hill, Catherine A., additional, Kuhn, Richard J., additional, and Perera, Rushika, additional

Published
2018
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18. Genomic insights into the Ixodes scapularis tick vector of Lyme disease

Author

Gulia-Nuss, Monika, Nuss, Andrew B., Meyer, Jason M., Sonenshine, Daniel E., Roe, R. Michael, Waterhouse, Robert M., Sattelle, David B., de la Fuente, Jose, Ribeiro, Jose M., Megy, Karine, Thimmapuram, Jyothi, Miller, Jason R., Walenz, Brian P., Koren, Sergey, Hostetler, Jessica B., Thiagarajan, Mathangi, Joardar, Vinita S., Hannick, Linda I., Bidwell, Shelby, Hammond, Martin P., Young, Sarah, Zeng, Qiandong, Abrudan, Jenica L., Almeida, Francisca C., Ayllon, Nieves, Bhide, Ketaki, Bissinger, Brooke W., Bonzon-Kulichenko, Elena, Buckingham, Steven D., Caffrey, Daniel R., Caimano, Melissa J., Croset, Vincent, Driscoll, Timothy, Gilbert, Don, Gillespie, Joseph J., Giraldo-Calderon, Gloria I., Grabowski, Jeffrey M., Jiang, David, Khalil, Sayed M. S., Kim, Donghun, Kocan, Katherine M., Koci, Juraj, Kuhn, Richard J., Kurtti, Timothy J., Lees, Kristin, Lang, Emma G., Kennedy, Ryan C., Kwon, Hyeogsun, Perera, Rushika, Qi, Yumin, Radolf, Justin D., Sakamoto, Joyce M., Sanchez-Gracia, Alejandro, Severo, Maiara S., Silverman, Neal, Simo, Ladislav, Tojo, Marta, Tornador, Cristian, Van Zee, Janice P., Vazquez, Jesus, Vieira, Filipe G., Villar, Margarita, Wespiser, Adam R., Yang, Yunlong, Zhu, Jiwei, Arensburger, Peter, Pietrantonio, Patricia V., Barker, Stephen C., Shao, Renfu, Zdobnov, Evgeny M., Hauser, Frank, Grimmelikhuijzen, Cornelis J. P., Park, Yoonseong, Rozas, Julio, Benton, Richard, Pedra, Joao H. F., Nelson, David R., Unger, Maria F., Tubio, Jose M. C., Tu, Zhijian Jake, Robertson, Hugh M., Shumway, Martin, Sutton, Granger, Wortman, Jennifer R., Lawson, Daniel, Wikel, Stephen K., Nene, Vishvanath M., Fraser, Claire M., Collins, Frank H., Birren, Bruce, Nelson, Karen E., Caler, Elisabet, Hill, Catherine A., Gulia-Nuss, Monika, Nuss, Andrew B., Meyer, Jason M., Sonenshine, Daniel E., Roe, R. Michael, Waterhouse, Robert M., Sattelle, David B., de la Fuente, Jose, Ribeiro, Jose M., Megy, Karine, Thimmapuram, Jyothi, Miller, Jason R., Walenz, Brian P., Koren, Sergey, Hostetler, Jessica B., Thiagarajan, Mathangi, Joardar, Vinita S., Hannick, Linda I., Bidwell, Shelby, Hammond, Martin P., Young, Sarah, Zeng, Qiandong, Abrudan, Jenica L., Almeida, Francisca C., Ayllon, Nieves, Bhide, Ketaki, Bissinger, Brooke W., Bonzon-Kulichenko, Elena, Buckingham, Steven D., Caffrey, Daniel R., Caimano, Melissa J., Croset, Vincent, Driscoll, Timothy, Gilbert, Don, Gillespie, Joseph J., Giraldo-Calderon, Gloria I., Grabowski, Jeffrey M., Jiang, David, Khalil, Sayed M. S., Kim, Donghun, Kocan, Katherine M., Koci, Juraj, Kuhn, Richard J., Kurtti, Timothy J., Lees, Kristin, Lang, Emma G., Kennedy, Ryan C., Kwon, Hyeogsun, Perera, Rushika, Qi, Yumin, Radolf, Justin D., Sakamoto, Joyce M., Sanchez-Gracia, Alejandro, Severo, Maiara S., Silverman, Neal, Simo, Ladislav, Tojo, Marta, Tornador, Cristian, Van Zee, Janice P., Vazquez, Jesus, Vieira, Filipe G., Villar, Margarita, Wespiser, Adam R., Yang, Yunlong, Zhu, Jiwei, Arensburger, Peter, Pietrantonio, Patricia V., Barker, Stephen C., Shao, Renfu, Zdobnov, Evgeny M., Hauser, Frank, Grimmelikhuijzen, Cornelis J. P., Park, Yoonseong, Rozas, Julio, Benton, Richard, Pedra, Joao H. F., Nelson, David R., Unger, Maria F., Tubio, Jose M. C., Tu, Zhijian Jake, Robertson, Hugh M., Shumway, Martin, Sutton, Granger, Wortman, Jennifer R., Lawson, Daniel, Wikel, Stephen K., Nene, Vishvanath M., Fraser, Claire M., Collins, Frank H., Birren, Bruce, Nelson, Karen E., Caler, Elisabet, and Hill, Catherine A.

Abstract

Ticks transmit more pathogens to humans and animals than any other arthropod. We describe the 2.1 Gbp nuclear genome of the tick, Ixodes scapularis (Say), which vectors pathogens that cause Lyme disease, human granulocytic anaplasmosis, babesiosis and other diseases. The large genome reflects accumulation of repetitive DNA, new lineages of retro-transposons, and gene architecture patterns resembling ancient metazoans rather than pancrustaceans. Annotation of scaffolds representing similar to 57% of the genome, reveals 20,486 protein-coding genes and expansions of gene families associated with tick-host interactions. We report insights from genome analyses into parasitic processes unique to ticks, including host 'questing', prolonged feeding, cuticle synthesis, blood meal concentration, novel methods of haemoglobin digestion, haem detoxification, vitellogenesis and prolonged off-host survival. We identify proteins associated with the agent of human granulocytic anaplasmosis, an emerging disease, and the encephalitis-causing Langat virus, and a population structure correlated to life-history traits and transmission of the Lyme disease agent.

Published
2016
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19. Genomic insights into the Ixodes scapularis tick vector of Lyme disease

Author

Biochemistry, Fralin Life Sciences Institute, Gulia-Nuss, Monika, Nuss, Andrew B., Meyer, Jason M., Sonenshine, Daniel E., Roe, R. Michael, Waterhouse, Robert M., Sattelle, David B., de la Fuente, Jose, Ribeiro, Jose M., Megy, Karine, Thimmapuram, Jyothi, Miller, Jason R., Walenz, Brian P., Koren, Sergey, Hostetler, Jessica B., Thiagarajan, Mathangi, Joardar, Vinita S., Hannick, Linda I., Bidwell, Shelby, Hammond, Martin P., Young, Sarah, Zeng, Qiandong, Abrudan, Jenica L., Almeida, Francisca C., Ayllon, Nieves, Bhide, Ketaki, Bissinger, Brooke W., Bonzon-Kulichenko, Elena, Buckingham, Steven D., Caffrey, Daniel R., Caimano, Melissa J., Croset, Vincent, Driscoll, Timothy, Gilbert, Don, Gillespie, Joseph J., Giraldo-Calderon, Gloria I., Grabowski, Jeffrey M., Jiang, David, Khalil, Sayed M. S., Kim, Donghun, Kocan, Katherine M., Koci, Juraj, Kuhn, Richard J., Kurtti, Timothy J., Lees, Kristin, Lang, Emma G., Kennedy, Ryan C., Kwon, Hyeogsun, Perera, Rushika, Qi, Yumin, Radolf, Justin D., Sakamoto, Joyce M., Sanchez-Gracia, Alejandro, Severo, Maiara S., Silverman, Neal, Simo, Ladislav, Tojo, Marta, Tornador, Cristian, Van Zee, Janice P., Vazquez, Jesus, Vieira, Filipe G., Villar, Margarita, Wespiser, Adam R., Yang, Yunlong, Zhu, Jiwei, Arensburger, Peter, Pietrantonio, Patricia V., Barker, Stephen C., Shao, Renfu, Zdobnov, Evgeny M., Hauser, Frank, Grimmelikhuijzen, Cornelis J. P., Park, Yoonseong, Rozas, Julio, Benton, Richard, Pedra, Joao H. F., Nelson, David R., Unger, Maria F., Tubio, Jose M. C., Tu, Zhijian Jake, Robertson, Hugh M., Shumway, Martin, Sutton, Granger, Wortman, Jennifer R., Lawson, Daniel, Wikel, Stephen K., Nene, Vishvanath M., Fraser, Claire M., Collins, Frank H., Birren, Bruce, Nelson, Karen E., Caler, Elisabet, Hill, Catherine A., Biochemistry, Fralin Life Sciences Institute, Gulia-Nuss, Monika, Nuss, Andrew B., Meyer, Jason M., Sonenshine, Daniel E., Roe, R. Michael, Waterhouse, Robert M., Sattelle, David B., de la Fuente, Jose, Ribeiro, Jose M., Megy, Karine, Thimmapuram, Jyothi, Miller, Jason R., Walenz, Brian P., Koren, Sergey, Hostetler, Jessica B., Thiagarajan, Mathangi, Joardar, Vinita S., Hannick, Linda I., Bidwell, Shelby, Hammond, Martin P., Young, Sarah, Zeng, Qiandong, Abrudan, Jenica L., Almeida, Francisca C., Ayllon, Nieves, Bhide, Ketaki, Bissinger, Brooke W., Bonzon-Kulichenko, Elena, Buckingham, Steven D., Caffrey, Daniel R., Caimano, Melissa J., Croset, Vincent, Driscoll, Timothy, Gilbert, Don, Gillespie, Joseph J., Giraldo-Calderon, Gloria I., Grabowski, Jeffrey M., Jiang, David, Khalil, Sayed M. S., Kim, Donghun, Kocan, Katherine M., Koci, Juraj, Kuhn, Richard J., Kurtti, Timothy J., Lees, Kristin, Lang, Emma G., Kennedy, Ryan C., Kwon, Hyeogsun, Perera, Rushika, Qi, Yumin, Radolf, Justin D., Sakamoto, Joyce M., Sanchez-Gracia, Alejandro, Severo, Maiara S., Silverman, Neal, Simo, Ladislav, Tojo, Marta, Tornador, Cristian, Van Zee, Janice P., Vazquez, Jesus, Vieira, Filipe G., Villar, Margarita, Wespiser, Adam R., Yang, Yunlong, Zhu, Jiwei, Arensburger, Peter, Pietrantonio, Patricia V., Barker, Stephen C., Shao, Renfu, Zdobnov, Evgeny M., Hauser, Frank, Grimmelikhuijzen, Cornelis J. P., Park, Yoonseong, Rozas, Julio, Benton, Richard, Pedra, Joao H. F., Nelson, David R., Unger, Maria F., Tubio, Jose M. C., Tu, Zhijian Jake, Robertson, Hugh M., Shumway, Martin, Sutton, Granger, Wortman, Jennifer R., Lawson, Daniel, Wikel, Stephen K., Nene, Vishvanath M., Fraser, Claire M., Collins, Frank H., Birren, Bruce, Nelson, Karen E., Caler, Elisabet, and Hill, Catherine A.

Abstract

Ticks transmit more pathogens to humans and animals than any other arthropod. We describe the 2.1 Gbp nuclear genome of the tick, Ixodes scapularis (Say), which vectors pathogens that cause Lyme disease, human granulocytic anaplasmosis, babesiosis and other diseases. The large genome reflects accumulation of repetitive DNA, new lineages of retro-transposons, and gene architecture patterns resembling ancient metazoans rather than pancrustaceans. Annotation of scaffolds representing similar to 57% of the genome, reveals 20,486 protein-coding genes and expansions of gene families associated with tick-host interactions. We report insights from genome analyses into parasitic processes unique to ticks, including host 'questing', prolonged feeding, cuticle synthesis, blood meal concentration, novel methods of haemoglobin digestion, haem detoxification, vitellogenesis and prolonged off-host survival. We identify proteins associated with the agent of human granulocytic anaplasmosis, an emerging disease, and the encephalitis-causing Langat virus, and a population structure correlated to life-history traits and transmission of the Lyme disease agent.

Published
2016

20. Genomic insights into the Ixodes scapularis tick vector of Lyme disease

Author

National Institute of Allergy and Infectious Diseases (US), National Institutes of Health (US), Department of Health and Human Services (US), Australian Research Council, Ministerio de Ciencia e Innovación (España), National Science Foundation (US), Xunta de Galicia, European Commission, Department of Agriculture (US), Texas AgriLife Research, European Research Council, Swiss National Science Foundation, Boehringer Ingelheim Fonds, Fundação para a Ciência e a Tecnologia (Portugal), Lundbeck Foundation, Gulia-Nuss, Monika, Nuss, Andrew B., Meyer, Jason M., Sonenshine, Daniel E., Roe, R. Michael, Waterhouse, Robert M., Sattelle, David B., Fuente, José de la, Ribeiro, Jose M., Megy, Karine, Thimmapuram, Jyothi, Miller, Jason R., Walenz, Brian P., Koren, Sergey, Hostetler, Jessica B., Thiagarajan, Mathangi, Joardar, Vinita S., Hannick, Linda I., Bidwell, Shelby, Hammond, Martin P., Young, Sarah, Zeng, Qiandong, Abrudan, Jenica L., Almeida, Francisca C., Ayllón, Nieves, Bhide, Ketaki, Bissinger, Brooke W., Bonzón-Kulichenko, Elena, Buckingham, Steven D., Caffrey, Daniel R., Caimano, Melissa J., Croset, Vincent, Driscoll, Timothy, Gilbert, Don, Gillespie, Joseph J., Giraldo-Calderón, Gloria I., Grabowski, Jeffrey M., Jiang, David, Khalil, Sayed M .S., Kim, Donghun, Kocan, Katherine M., Koči, Juraj, Kuhn, Richard J., Kurtti, Timothy J., Lees, Kristin, Lang, Emma G., Kennedy, Ryan C., Kwon, Hyeogsun, Perera, Rushika, Qi, Yumin, Radolf, Justin D., Sakamoto, Joyce M., Sánchez-Gracia, Alejandro, Severo, Maiara S., Silverman, Neal, Šimo, Ladislav, Tojo, Marta, Tornador, Cristian, Van Zee, Janice P., Vázquez, Jesús, Vieira, Filipe G., Villar, Margarita, Wespiser, Adam R., Yang, Yunlong, Zhu, Jiwei, Arensburger, Peter, Pietrantonio, Patricia V., Barker, Stephen C., Shao, Renfu, Zdobnov, Evgeny M., Hauser, Frank, Grimmelikhuijzen, Cornelis J. P., Park, Yoonseong, Rozas, Julio, Benton, Richard, Pedra, Joao H. F., Nelson, David R., Unger, Maria F., Tubio, Jose M. C., Tu, Zhijian, Robertson, Hugh M., Shumway, Martin, Sutton, Granger, Wortman, Jennifer R., Lawson, Daniel, Wikel, Stephen K., Nene, Vishvanath M., Fraser, Claire M., Collins, Frank H., Birren, Bruce, Nelson, Karen E., Caler, Elisabet, Hill, Catherine A., National Institute of Allergy and Infectious Diseases (US), National Institutes of Health (US), Department of Health and Human Services (US), Australian Research Council, Ministerio de Ciencia e Innovación (España), National Science Foundation (US), Xunta de Galicia, European Commission, Department of Agriculture (US), Texas AgriLife Research, European Research Council, Swiss National Science Foundation, Boehringer Ingelheim Fonds, Fundação para a Ciência e a Tecnologia (Portugal), Lundbeck Foundation, Gulia-Nuss, Monika, Nuss, Andrew B., Meyer, Jason M., Sonenshine, Daniel E., Roe, R. Michael, Waterhouse, Robert M., Sattelle, David B., Fuente, José de la, Ribeiro, Jose M., Megy, Karine, Thimmapuram, Jyothi, Miller, Jason R., Walenz, Brian P., Koren, Sergey, Hostetler, Jessica B., Thiagarajan, Mathangi, Joardar, Vinita S., Hannick, Linda I., Bidwell, Shelby, Hammond, Martin P., Young, Sarah, Zeng, Qiandong, Abrudan, Jenica L., Almeida, Francisca C., Ayllón, Nieves, Bhide, Ketaki, Bissinger, Brooke W., Bonzón-Kulichenko, Elena, Buckingham, Steven D., Caffrey, Daniel R., Caimano, Melissa J., Croset, Vincent, Driscoll, Timothy, Gilbert, Don, Gillespie, Joseph J., Giraldo-Calderón, Gloria I., Grabowski, Jeffrey M., Jiang, David, Khalil, Sayed M .S., Kim, Donghun, Kocan, Katherine M., Koči, Juraj, Kuhn, Richard J., Kurtti, Timothy J., Lees, Kristin, Lang, Emma G., Kennedy, Ryan C., Kwon, Hyeogsun, Perera, Rushika, Qi, Yumin, Radolf, Justin D., Sakamoto, Joyce M., Sánchez-Gracia, Alejandro, Severo, Maiara S., Silverman, Neal, Šimo, Ladislav, Tojo, Marta, Tornador, Cristian, Van Zee, Janice P., Vázquez, Jesús, Vieira, Filipe G., Villar, Margarita, Wespiser, Adam R., Yang, Yunlong, Zhu, Jiwei, Arensburger, Peter, Pietrantonio, Patricia V., Barker, Stephen C., Shao, Renfu, Zdobnov, Evgeny M., Hauser, Frank, Grimmelikhuijzen, Cornelis J. P., Park, Yoonseong, Rozas, Julio, Benton, Richard, Pedra, Joao H. F., Nelson, David R., Unger, Maria F., Tubio, Jose M. C., Tu, Zhijian, Robertson, Hugh M., Shumway, Martin, Sutton, Granger, Wortman, Jennifer R., Lawson, Daniel, Wikel, Stephen K., Nene, Vishvanath M., Fraser, Claire M., Collins, Frank H., Birren, Bruce, Nelson, Karen E., Caler, Elisabet, and Hill, Catherine A.

Abstract

Ticks transmit more pathogens to humans and animals than any other arthropod. We describe the 2.1 Gbp nuclear genome of the tick, Ixodes scapularis (Say), which vectors pathogens that cause Lyme disease, human granulocytic anaplasmosis, babesiosis and other diseases. The large genome reflects accumulation of repetitive DNA, new lineages of retro-transposons, and gene architecture patterns resembling ancient metazoans rather than pancrustaceans. Annotation of scaffolds representing ∼57% of the genome, reveals 20,486 protein-coding genes and expansions of gene families associated with tick–host interactions. We report insights from genome analyses into parasitic processes unique to ticks, including host ‘questing’, prolonged feeding, cuticle synthesis, blood meal concentration, novel methods of haemoglobin digestion, haem detoxification, vitellogenesis and prolonged off-host survival. We identify proteins associated with the agent of human granulocytic anaplasmosis, an emerging disease, and the encephalitis-causing Langat virus, and a population structure correlated to life-history traits and transmission of the Lyme disease agent.

Published
2016

21. Changes in the Proteome of Langat-Infected Ixodes scapularis ISE6 Cells: Metabolic Pathways Associated with Flavivirus Infection

Author

Grabowski, Jeffrey M., Perera, Rushika, Roumani, Ali M., Hedrick, Victoria E., Inerowicz, Halina Dorota, Hill, Catherine A., Kuhn, Richard, Grabowski, Jeffrey M., Perera, Rushika, Roumani, Ali M., Hedrick, Victoria E., Inerowicz, Halina Dorota, Hill, Catherine A., and Kuhn, Richard

Abstract

Background: Ticks (Family Ixodidae) transmit a variety of disease causing agents to humans and animals. The tick-borne flaviviruses (TBFs; family Flaviviridae) are a complex of viruses, many of which cause encephalitis and hemorrhagic fever, and represent global threats to human health and biosecurity. Pathogenesis has been well studied in human and animal disease models. Equivalent analyses of tick-flavivirus interactions are limited and represent an area of study that could reveal novel approaches for TBF control. Methodology/Principal Findings: High resolution LC-MS/MS was used to analyze the proteome of Ixodes scapularis (Lyme disease tick) embryonic ISE6 cells following infection with Langat virus (LGTV) and identify proteins associated with viral infection and replication. Maximal LGTV infection of cells and determination of peak release of infectious virus, was observed at 36 hours post infection (hpi). Proteins were extracted from ISE6 cells treated with LGTV and non-infectious (UV inactivated) LGTV at 36 hpi and analyzed by mass spectrometry. The Omics Discovery Pipeline (ODP) identified thousands of MS peaks. Protein homology searches against the I. scapularis IscaW1 genome assembly identified a total of 486 proteins that were subsequently assigned to putative functional pathways using searches against the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. 266 proteins were differentially expressed following LGTV infection relative to non-infected (mock) cells. Of these, 68 proteins exhibited increased expression and 198 proteins had decreased expression. The majority of the former were classified in the KEGG pathways: “translation”, “amino acid metabolism”, and “protein folding/sorting/degradation”. Finally, Trichostatin A and Oligomycin A increased and decreased LGTV replication in vitro in ISE6 cells, respectively. Conclusions/Significance: Proteomic analyses revealed ISE6 proteins that were differentially expressed at the peak of LGTV replication

Published
2016

23. Genomic insights into the Ixodes scapularis tick vector of Lyme disease

Author

Gulia-Nuss, Monika, primary, Nuss, Andrew B., additional, Meyer, Jason M., additional, Sonenshine, Daniel E., additional, Roe, R. Michael, additional, Waterhouse, Robert M., additional, Sattelle, David B., additional, de la Fuente, José, additional, Ribeiro, Jose M., additional, Megy, Karine, additional, Thimmapuram, Jyothi, additional, Miller, Jason R., additional, Walenz, Brian P., additional, Koren, Sergey, additional, Hostetler, Jessica B., additional, Thiagarajan, Mathangi, additional, Joardar, Vinita S., additional, Hannick, Linda I., additional, Bidwell, Shelby, additional, Hammond, Martin P., additional, Young, Sarah, additional, Zeng, Qiandong, additional, Abrudan, Jenica L., additional, Almeida, Francisca C., additional, Ayllón, Nieves, additional, Bhide, Ketaki, additional, Bissinger, Brooke W., additional, Bonzon-Kulichenko, Elena, additional, Buckingham, Steven D., additional, Caffrey, Daniel R., additional, Caimano, Melissa J., additional, Croset, Vincent, additional, Driscoll, Timothy, additional, Gilbert, Don, additional, Gillespie, Joseph J., additional, Giraldo-Calderón, Gloria I., additional, Grabowski, Jeffrey M., additional, Jiang, David, additional, Khalil, Sayed M. S., additional, Kim, Donghun, additional, Kocan, Katherine M., additional, Koči, Juraj, additional, Kuhn, Richard J., additional, Kurtti, Timothy J., additional, Lees, Kristin, additional, Lang, Emma G., additional, Kennedy, Ryan C., additional, Kwon, Hyeogsun, additional, Perera, Rushika, additional, Qi, Yumin, additional, Radolf, Justin D., additional, Sakamoto, Joyce M., additional, Sánchez-Gracia, Alejandro, additional, Severo, Maiara S., additional, Silverman, Neal, additional, Šimo, Ladislav, additional, Tojo, Marta, additional, Tornador, Cristian, additional, Van Zee, Janice P., additional, Vázquez, Jesús, additional, Vieira, Filipe G., additional, Villar, Margarita, additional, Wespiser, Adam R., additional, Yang, Yunlong, additional, Zhu, Jiwei, additional, Arensburger, Peter, additional, Pietrantonio, Patricia V., additional, Barker, Stephen C., additional, Shao, Renfu, additional, Zdobnov, Evgeny M., additional, Hauser, Frank, additional, Grimmelikhuijzen, Cornelis J. P., additional, Park, Yoonseong, additional, Rozas, Julio, additional, Benton, Richard, additional, Pedra, Joao H. F., additional, Nelson, David R., additional, Unger, Maria F., additional, Tubio, Jose M. C., additional, Tu, Zhijian, additional, Robertson, Hugh M., additional, Shumway, Martin, additional, Sutton, Granger, additional, Wortman, Jennifer R., additional, Lawson, Daniel, additional, Wikel, Stephen K., additional, Nene, Vishvanath M., additional, Fraser, Claire M., additional, Collins, Frank H., additional, Birren, Bruce, additional, Nelson, Karen E., additional, Caler, Elisabet, additional, and Hill, Catherine A., additional

Published
2016
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25. RNAi reveals proteins for metabolism and protein processing associated with Langat virus infection in Ixodes scapularis (blacklegged tick) ISE6 cells.

Author

Grabowski, Jeffrey M., Gulia-Nuss, Monika, Kuhn, Richard J., and Hill, Catherine A.

Subjects
*IXODES scapularis, *TICK-borne diseases, *FLAVIVIRUSES, *ENCEPHALITIS, *INFECTIOUS disease transmission
Abstract

Background: Tick-borne flaviviruses (TBFs) cause thousands of human cases of encephalitis worldwide each year, with some TBF infections progressing to hemorrhagic fever. TBFs are of medical and veterinary importance and strategies to reduce flavivirus transmission by the tick vector may have significant application. Analyses of the proteome of ISE6 cells derived from the black legged tick, Ixodes scapularis infected with the TBF, Langat virus (LGTV), have provided insights into proteins and cellular processes involved with LGTV infection. Methods: RNA interference (RNAi)-induced knockdown of transcripts was used to investigate the role of ten tick proteins in the LGTV infection cycle in ISE6 cells. LGTV-infected cells were separately transfected with dsRNA corresponding to each gene of interest and the effect on LGTV genome replication and release of infectious virus was assessed by RT-qPCR and plaque assays, respectively. Results: RNAi-induced knockdown of transcripts for two enzymes that likely function in amino acid, carbohydrate, lipid, terpenoid/polykeytide and vitamin metabolism, and a transcript for one protein of unknown function were associated with decreased replication of the LGTV genome and release of infectious virus from cells. The knockdown of transcripts for five enzymes predicted to function in metabolism, a protein likely associated with folding, sorting and degradation, and a protein of unknown function was associated with a decrease only in the amount of infectious LGTV released from cells. Conclusions: These data suggest tick proteins potentially associated with metabolism and protein processing may be involved in LGTV infection of ISE6 cells. Our study provides information to begin to elucidate the function of these proteins and identify targets for the development of new interventions aimed at controlling the transmission of TBFs. [ABSTRACT FROM AUTHOR]

Published
2017
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27. Dissecting Flavivirus Biology in Salivary Gland Cultures from Fed and Unfed Ixodes scapularis(Black-Legged Tick)

Author

Grabowski, Jeffrey M., Nilsson, Olof R., Fischer, Elizabeth R., Long, Dan, Offerdahl, Danielle K., Park, Yoonseong, Scott, Dana P., and Bloom, Marshall E.

Abstract

Tick-borne flaviviruses (TBFVs) are responsible for more than 15,000 human disease cases each year, and Powassan virus lineage 2 (POWV-L2) deer tick virus has been a reemerging threat in North America over the past 20 years. Rapid transmission of TBFVs in particular emphasizes the importance of preventing tick bites, the difficulty in developing countermeasures to prevent transmission, and the importance of understanding TBFV infection in tick salivary glands (SGs). Tick blood feeding is responsible for phenomenal physiological changes and is associated with changes in TBFV multiplication within the tick and in SGs. Using SG cultures from Ixodes scapularisfemale ticks, the primary aims of this study were to identify cellular localization of virus-like particles in acini of infected SGs from fed and unfed ticks, localization of TBFV infection in infected SGs from fed ticks, and a tick transcript (with associated metabolic function) involved in POWV-L2 infection in SG cultures.

Published
2019
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28. Flavivirus Infection of Ixodes scapularis(Black-Legged Tick) Ex VivoOrganotypic Cultures and Applications for Disease Control

Author

Grabowski, Jeffrey M., Tsetsarkin, Konstantin A., Long, Dan, Scott, Dana P., Rosenke, Rebecca, Schwan, Tom G., Mlera, Luwanika, Offerdahl, Danielle K., Pletnev, Alexander G., and Bloom, Marshall E.

Abstract

ABSTRACTIxodes scapularisticks transmit many infectious agents that cause disease, including tick-borne flaviviruses (TBFVs). TBFV infections cause thousands of human encephalitis cases worldwide annually. In the United States, human TBFV infections with Powassan virus (POWV) are increasing and have a fatality rate of 10 to 30%. Additionally, Langat virus (LGTV) is a TBFV of low neurovirulence and is used as a model TBFV. TBFV replication and dissemination within I. scapularisorgans are poorly characterized, and a deeper understanding of virus biology in this vector may inform effective countermeasures to reduce TBFV transmission. Here, we describe short-term, I. scapularisorgan culture models of TBFV infection. Ex vivoorgans were metabolically active for 9 to 10 days and were permissive to LGTV and POWV replication. Imaging and videography demonstrated replication and spread of green fluorescent protein-expressing LGTV in the organs. Immunohistochemical staining confirmed LGTV envelope and POWV protein synthesis within the infected organs. LGTV- and POWV-infected organs produced infectious LGTV and POWV; thus, the ex vivocultures were suitable for study of virus replication in individual organs. LGTV- and POWV-infected midgut and salivary glands were subjected to double-stranded RNA (dsRNA) transfection with dsRNA to the LGTV 3′ untranslated region (UTR), which reduced infectious LGTV and POWV replication, providing a proof-of-concept use of RNA interference in I. scapularisorgan cultures to study the effects on TBFV replication. The results contribute important information on TBFV localization within ex vivo I. scapularisorgans and provide a significant translational tool for evaluating recombinant, live vaccine candidates and potential tick transcripts and proteins for possible therapeutic use and vaccine development to reduce TBFV transmission.IMPORTANCETick-borne flavivirus (TBFV) infections cause neurological and/or hemorrhagic disease in humans worldwide. There are currently no licensed therapeutics or vaccines against Powassan virus (POWV), the only TBFV known to circulate in North America. Evaluating tick vector targets for antitick vaccines directed at reducing TBFV infection within the arthropod vector is a critical step in identifying efficient approaches to controlling TBFV transmission. This study characterized infection of female Ixodes scapularistick organ cultures of midgut, salivary glands, and synganglion with the low-neurovirulence Langat virus (LGTV) and the more pathogenic POWV. Cell types of specific organs were susceptible to TBFV infection, and a difference in LGTV and POWV replication was noted in TBFV-infected organs. This tick organ culture model of TBFV infection will be useful for various applications, such as screening of tick endogenous dsRNA corresponding to potential control targets within midgut and salivary glands to confirm restriction of TBFV infection.

Published
2017
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29. Impact of Dengue Virus Infection on Global Metabolic Alterations in the Aedes aegyptiMosquito Vector

Author

Chotiwan, Nunya, Sanchez-Vargus, Irma, Grabowski, Jeffrey M., Hopf-jannasch, Amber, Hedrick, Victoria, Gough, Erik, Nakayasu, Ernesto, Sirohi, Devika, Hill, Catherine A., Kuhn, Richard J., and Perera, Rushika

Abstract

Aedes aegyptimosquitoes are the primary vectors transmitting dengue virus (DENV), one of the most aggressive re-emerging pathogens worldwide causing more than 390 million infections per year. The spread of the virus is greatly dependent upon successful replication within both the human host and mosquito vector. Much effort has been placed in understanding the dynamics of virus transmission and replication in both organisms, but little is known about the global impact of DENV on metabolic pathways. Previous studies have demonstrated perturbations in human and Aedes albopictuscellular metabolic environments during DENV infection. Some of these perturbations include increasing the production of membranous lipids that had the capability to induce membrane curvature and permeability, as well as visibly altering both human and mosquito intracellular membrane architecture to support DENV replication. In this study, we have explored metabolic changes in Aedes aegyptimidgut and salivary glands upon DENV (serotype 2) infection. We have found several significant fluctuations in the lipid and metabolite repertoire from infected tissues compared to uninfected controls, including differential expression of molecules that function as membrane building blocks, bioactive messengers, energy storage and intermediates in lipid biosynthesis and lipolysis pathways. These results and their relevance to dengue virus infection of its mosquito vector will be discussed.

Published
2015
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30. Characterization of the proteome of cytoplasmic lipid droplets in mouse enterocytes after a dietary fat challenge.

Author

D'Aquila T, Sirohi D, Grabowski JM, Hedrick VE, Paul LN, Greenberg AS, Kuhn RJ, and Buhman KK

Subjects
Animals, Apolipoproteins A metabolism, Carrier Proteins metabolism, Coenzyme A Ligases metabolism, Enterocytes ultrastructure, Lipid Droplets ultrastructure, Lipid Metabolism, Male, Metabolic Networks and Pathways, Mice, Mice, Inbred C57BL, Microscopy, Electron, Transmission, Models, Biological, Perilipin-3, Triglycerides metabolism, Dietary Fats administration & dosage, Enterocytes metabolism, Lipid Droplets metabolism, Proteome metabolism
Abstract

Dietary fat absorption by the small intestine is a multistep process that regulates the uptake and delivery of essential nutrients and energy. One step of this process is the temporary storage of dietary fat in cytoplasmic lipid droplets (CLDs). The storage and mobilization of dietary fat is thought to be regulated by proteins that associate with the CLD; however, mechanistic details of this process are currently unknown. In this study we analyzed the proteome of CLDs isolated from enterocytes harvested from the small intestine of mice following a dietary fat challenge. In this analysis we identified 181 proteins associated with the CLD fraction, of which 37 are associated with known lipid related metabolic pathways. We confirmed the localization of several of these proteins on or around the CLD through confocal and electron microscopy, including perilipin 3, apolipoprotein A-IV, and acyl-CoA synthetase long-chain family member 5. The identification of the enterocyte CLD proteome provides new insight into potential regulators of CLD metabolism and the process of dietary fat absorption.

Published
2015
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