Cargando…

HCV NS5A dimer interface residues regulate HCV replication by controlling its self-interaction, hyperphosphorylation, subcellular localization and interaction with cyclophilin A

The HCV NS5A protein plays multiple roles during viral replication, including viral genome replication and virus particle assembly. The crystal structures of the NS5A N-terminal domain indicated the potential existence of the NS5A dimers formed via at least two or more distinct dimeric interfaces. H...

Descripción completa

Detalles Bibliográficos
Autores principales: Shanmugam, Saravanabalaji, Nichols, Alyssa K., Saravanabalaji, Dhanaranjani, Welsch, Christoph, Yi, MinKyung
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072203/
https://www.ncbi.nlm.nih.gov/pubmed/30036383
http://dx.doi.org/10.1371/journal.ppat.1007177
_version_ 1783343992890458112
author Shanmugam, Saravanabalaji
Nichols, Alyssa K.
Saravanabalaji, Dhanaranjani
Welsch, Christoph
Yi, MinKyung
author_facet Shanmugam, Saravanabalaji
Nichols, Alyssa K.
Saravanabalaji, Dhanaranjani
Welsch, Christoph
Yi, MinKyung
author_sort Shanmugam, Saravanabalaji
collection PubMed
description The HCV NS5A protein plays multiple roles during viral replication, including viral genome replication and virus particle assembly. The crystal structures of the NS5A N-terminal domain indicated the potential existence of the NS5A dimers formed via at least two or more distinct dimeric interfaces. However, it is unknown whether these different forms of NS5A dimers are involved in its numerous functions. To address this question, we mutated the residues lining the two different NS5A dimer interfaces and determined their effects on NS5A self-interaction, NS5A-cyclophilin A (CypA) interaction, HCV RNA replication and infectious virus production. We found that the mutations targeting either of two dimeric interfaces disrupted the NS5A self-interaction in cells. The NS5A dimer-interrupting mutations also inhibited both viral RNA replication and infectious virus production with some genotypic differences. We also determined that reduced NS5A self-interaction was associated with altered NS5A-CypA interaction, NS5A hyperphosphorylation and NS5A subcellular localization, providing the mechanistic bases for the role of NS5A self-interaction in multiple steps of HCV replication. The NS5A oligomers formed via different interfaces are likely its functional form, since the residues at two different dimeric interfaces played similar roles in different aspects of NS5A functions and, consequently, HCV replication. In conclusion, this study provides novel insight into the functional significance of NS5A self-interaction in different steps of the HCV replication, potentially, in the form of oligomers formed via multiple dimeric interfaces.
format Online
Article
Text
id pubmed-6072203
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-60722032018-08-16 HCV NS5A dimer interface residues regulate HCV replication by controlling its self-interaction, hyperphosphorylation, subcellular localization and interaction with cyclophilin A Shanmugam, Saravanabalaji Nichols, Alyssa K. Saravanabalaji, Dhanaranjani Welsch, Christoph Yi, MinKyung PLoS Pathog Research Article The HCV NS5A protein plays multiple roles during viral replication, including viral genome replication and virus particle assembly. The crystal structures of the NS5A N-terminal domain indicated the potential existence of the NS5A dimers formed via at least two or more distinct dimeric interfaces. However, it is unknown whether these different forms of NS5A dimers are involved in its numerous functions. To address this question, we mutated the residues lining the two different NS5A dimer interfaces and determined their effects on NS5A self-interaction, NS5A-cyclophilin A (CypA) interaction, HCV RNA replication and infectious virus production. We found that the mutations targeting either of two dimeric interfaces disrupted the NS5A self-interaction in cells. The NS5A dimer-interrupting mutations also inhibited both viral RNA replication and infectious virus production with some genotypic differences. We also determined that reduced NS5A self-interaction was associated with altered NS5A-CypA interaction, NS5A hyperphosphorylation and NS5A subcellular localization, providing the mechanistic bases for the role of NS5A self-interaction in multiple steps of HCV replication. The NS5A oligomers formed via different interfaces are likely its functional form, since the residues at two different dimeric interfaces played similar roles in different aspects of NS5A functions and, consequently, HCV replication. In conclusion, this study provides novel insight into the functional significance of NS5A self-interaction in different steps of the HCV replication, potentially, in the form of oligomers formed via multiple dimeric interfaces. Public Library of Science 2018-07-23 /pmc/articles/PMC6072203/ /pubmed/30036383 http://dx.doi.org/10.1371/journal.ppat.1007177 Text en © 2018 Shanmugam 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
Shanmugam, Saravanabalaji
Nichols, Alyssa K.
Saravanabalaji, Dhanaranjani
Welsch, Christoph
Yi, MinKyung
HCV NS5A dimer interface residues regulate HCV replication by controlling its self-interaction, hyperphosphorylation, subcellular localization and interaction with cyclophilin A
title HCV NS5A dimer interface residues regulate HCV replication by controlling its self-interaction, hyperphosphorylation, subcellular localization and interaction with cyclophilin A
title_full HCV NS5A dimer interface residues regulate HCV replication by controlling its self-interaction, hyperphosphorylation, subcellular localization and interaction with cyclophilin A
title_fullStr HCV NS5A dimer interface residues regulate HCV replication by controlling its self-interaction, hyperphosphorylation, subcellular localization and interaction with cyclophilin A
title_full_unstemmed HCV NS5A dimer interface residues regulate HCV replication by controlling its self-interaction, hyperphosphorylation, subcellular localization and interaction with cyclophilin A
title_short HCV NS5A dimer interface residues regulate HCV replication by controlling its self-interaction, hyperphosphorylation, subcellular localization and interaction with cyclophilin A
title_sort hcv ns5a dimer interface residues regulate hcv replication by controlling its self-interaction, hyperphosphorylation, subcellular localization and interaction with cyclophilin a
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072203/
https://www.ncbi.nlm.nih.gov/pubmed/30036383
http://dx.doi.org/10.1371/journal.ppat.1007177
work_keys_str_mv AT shanmugamsaravanabalaji hcvns5adimerinterfaceresiduesregulatehcvreplicationbycontrollingitsselfinteractionhyperphosphorylationsubcellularlocalizationandinteractionwithcyclophilina
AT nicholsalyssak hcvns5adimerinterfaceresiduesregulatehcvreplicationbycontrollingitsselfinteractionhyperphosphorylationsubcellularlocalizationandinteractionwithcyclophilina
AT saravanabalajidhanaranjani hcvns5adimerinterfaceresiduesregulatehcvreplicationbycontrollingitsselfinteractionhyperphosphorylationsubcellularlocalizationandinteractionwithcyclophilina
AT welschchristoph hcvns5adimerinterfaceresiduesregulatehcvreplicationbycontrollingitsselfinteractionhyperphosphorylationsubcellularlocalizationandinteractionwithcyclophilina
AT yiminkyung hcvns5adimerinterfaceresiduesregulatehcvreplicationbycontrollingitsselfinteractionhyperphosphorylationsubcellularlocalizationandinteractionwithcyclophilina