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Species A rotavirus NSP3 acquires its translation inhibitory function prior to stable dimer formation
Species A rotavirus non-structural protein 3 (NSP3) is a translational regulator that inhibits or, under some conditions, enhances host cell translation. NSP3 binds to the translation initiation factor eIF4G1 and evicts poly-(A) binding protein (PABP) from eIF4G1, thus inhibiting translation of poly...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Public Library of Science
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5524322/ https://www.ncbi.nlm.nih.gov/pubmed/28738064 http://dx.doi.org/10.1371/journal.pone.0181871 |
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author | Contreras-Treviño, Hugo I. Reyna-Rosas, Edgar León-Rodríguez, Renato Ruiz-Ordaz, Blanca H. Dinkova, Tzvetanka D. Cevallos, Ana M. Padilla-Noriega, Luis |
author_facet | Contreras-Treviño, Hugo I. Reyna-Rosas, Edgar León-Rodríguez, Renato Ruiz-Ordaz, Blanca H. Dinkova, Tzvetanka D. Cevallos, Ana M. Padilla-Noriega, Luis |
author_sort | Contreras-Treviño, Hugo I. |
collection | PubMed |
description | Species A rotavirus non-structural protein 3 (NSP3) is a translational regulator that inhibits or, under some conditions, enhances host cell translation. NSP3 binds to the translation initiation factor eIF4G1 and evicts poly-(A) binding protein (PABP) from eIF4G1, thus inhibiting translation of polyadenylated mRNAs, presumably by disrupting the effect of PABP bound to their 3’-ends. NSP3 has a long coiled-coil region involved in dimerization that includes a chaperone Hsp90-binding domain (HS90BD). We aimed to study the role in NSP3 dimerization of a segment of the coiled-coil region adjoining the HS90BD. We used a vaccinia virus system to express NSP3 with point mutations in conserved amino acids in the coiled-coil region and determined the effects of these mutations on translation by metabolic labeling of proteins as well as on accumulation of stable NSP3 dimers by non-dissociating Western blot, a method that separates stable NSP3 dimers from the monomer/dimerization intermediate forms of the protein. Four of five mutations reduced the total yield of NSP3 and the formation of stable dimers (W170A, K171E, R173E and R187E:K191E), whereas one mutation had the opposite effects (Y192A). Treatment with the proteasome inhibitor MG132 revealed that stable NSP3 dimers and monomers/dimerization intermediates are susceptible to proteasome degradation. Surprisingly, mutants severely impaired in the formation of stable dimers were still able to inhibit host cell translation, suggesting that NSP3 dimerization intermediates are functional. Our results demonstrate that rotavirus NSP3 acquires its function prior to stable dimer formation and remain as a proteasome target throughout dimerization. |
format | Online Article Text |
id | pubmed-5524322 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-55243222017-08-07 Species A rotavirus NSP3 acquires its translation inhibitory function prior to stable dimer formation Contreras-Treviño, Hugo I. Reyna-Rosas, Edgar León-Rodríguez, Renato Ruiz-Ordaz, Blanca H. Dinkova, Tzvetanka D. Cevallos, Ana M. Padilla-Noriega, Luis PLoS One Research Article Species A rotavirus non-structural protein 3 (NSP3) is a translational regulator that inhibits or, under some conditions, enhances host cell translation. NSP3 binds to the translation initiation factor eIF4G1 and evicts poly-(A) binding protein (PABP) from eIF4G1, thus inhibiting translation of polyadenylated mRNAs, presumably by disrupting the effect of PABP bound to their 3’-ends. NSP3 has a long coiled-coil region involved in dimerization that includes a chaperone Hsp90-binding domain (HS90BD). We aimed to study the role in NSP3 dimerization of a segment of the coiled-coil region adjoining the HS90BD. We used a vaccinia virus system to express NSP3 with point mutations in conserved amino acids in the coiled-coil region and determined the effects of these mutations on translation by metabolic labeling of proteins as well as on accumulation of stable NSP3 dimers by non-dissociating Western blot, a method that separates stable NSP3 dimers from the monomer/dimerization intermediate forms of the protein. Four of five mutations reduced the total yield of NSP3 and the formation of stable dimers (W170A, K171E, R173E and R187E:K191E), whereas one mutation had the opposite effects (Y192A). Treatment with the proteasome inhibitor MG132 revealed that stable NSP3 dimers and monomers/dimerization intermediates are susceptible to proteasome degradation. Surprisingly, mutants severely impaired in the formation of stable dimers were still able to inhibit host cell translation, suggesting that NSP3 dimerization intermediates are functional. Our results demonstrate that rotavirus NSP3 acquires its function prior to stable dimer formation and remain as a proteasome target throughout dimerization. Public Library of Science 2017-07-24 /pmc/articles/PMC5524322/ /pubmed/28738064 http://dx.doi.org/10.1371/journal.pone.0181871 Text en © 2017 Contreras-Treviño et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Contreras-Treviño, Hugo I. Reyna-Rosas, Edgar León-Rodríguez, Renato Ruiz-Ordaz, Blanca H. Dinkova, Tzvetanka D. Cevallos, Ana M. Padilla-Noriega, Luis Species A rotavirus NSP3 acquires its translation inhibitory function prior to stable dimer formation |
title | Species A rotavirus NSP3 acquires its translation inhibitory function prior to stable dimer formation |
title_full | Species A rotavirus NSP3 acquires its translation inhibitory function prior to stable dimer formation |
title_fullStr | Species A rotavirus NSP3 acquires its translation inhibitory function prior to stable dimer formation |
title_full_unstemmed | Species A rotavirus NSP3 acquires its translation inhibitory function prior to stable dimer formation |
title_short | Species A rotavirus NSP3 acquires its translation inhibitory function prior to stable dimer formation |
title_sort | species a rotavirus nsp3 acquires its translation inhibitory function prior to stable dimer formation |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5524322/ https://www.ncbi.nlm.nih.gov/pubmed/28738064 http://dx.doi.org/10.1371/journal.pone.0181871 |
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