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Nonstructural NS5A Protein Regulates LIM and SH3 Domain Protein 1 to Promote Hepatitis C Virus Propagation

Hepatitis C virus (HCV) propagation is highly dependent on cellular proteins. To identify the host factors involved in HCV propagation, we previously performed protein microarray assays and identified the LIM and SH3 domain protein 1 (LASP-1) as an HCV NS5A-interacting partner. LASP-1 plays an impor...

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Autores principales: Choi, Jae-Woong, Kim, Jong-Wook, Nguyen, Lap P., Nguyen, Huu C., Park, Eun-Mee, Choi, Dong Hwa, Han, Kang Min, Kang, Sang Min, Tark, Dongseob, Lim, Yun-Sook, Hwang, Soon B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Korean Society for Molecular and Cellular Biology 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7264479/
https://www.ncbi.nlm.nih.gov/pubmed/32344996
http://dx.doi.org/10.14348/molcells.2020.0018
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author Choi, Jae-Woong
Kim, Jong-Wook
Nguyen, Lap P.
Nguyen, Huu C.
Park, Eun-Mee
Choi, Dong Hwa
Han, Kang Min
Kang, Sang Min
Tark, Dongseob
Lim, Yun-Sook
Hwang, Soon B.
author_facet Choi, Jae-Woong
Kim, Jong-Wook
Nguyen, Lap P.
Nguyen, Huu C.
Park, Eun-Mee
Choi, Dong Hwa
Han, Kang Min
Kang, Sang Min
Tark, Dongseob
Lim, Yun-Sook
Hwang, Soon B.
author_sort Choi, Jae-Woong
collection PubMed
description Hepatitis C virus (HCV) propagation is highly dependent on cellular proteins. To identify the host factors involved in HCV propagation, we previously performed protein microarray assays and identified the LIM and SH3 domain protein 1 (LASP-1) as an HCV NS5A-interacting partner. LASP-1 plays an important role in the regulation of cell proliferation, migration, and protein-protein interactions. Alteration of LASP-1 expression has been implicated in hepatocellular carcinoma. However, the functional involvement of LASP1 in HCV propagation and HCV-induced pathogenesis has not been elucidated. Here, we first verified the protein interaction of NS5A and LASP-1 by both in vitro pulldown and coimmunoprecipitation assays. We further showed that NS5A and LASP-1 were colocalized in the cytoplasm of HCV infected cells. NS5A interacted with LASP-1 through the proline motif in domain I of NS5A and the tryptophan residue in the SH3 domain of LASP-1. Knockdown of LASP-1 increased HCV replication in both HCV-infected cells and HCV subgenomic replicon cells. LASP-1 negatively regulated viral propagation and thereby overexpression of LASP-1 decreased HCV replication. Moreover, HCV propagation was decreased by wild-type LASP-1 but not by an NS5A binding-defective mutant of LASP-1. We further demonstrated that LASP-1 was involved in the replication stage of the HCV life cycle. Importantly, LASP-1 expression levels were increased in persistently infected cells with HCV. These data suggest that HCV modulates LASP-1 via NS5A in order to regulate virion levels and maintain a persistent infection.
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spelling pubmed-72644792020-06-10 Nonstructural NS5A Protein Regulates LIM and SH3 Domain Protein 1 to Promote Hepatitis C Virus Propagation Choi, Jae-Woong Kim, Jong-Wook Nguyen, Lap P. Nguyen, Huu C. Park, Eun-Mee Choi, Dong Hwa Han, Kang Min Kang, Sang Min Tark, Dongseob Lim, Yun-Sook Hwang, Soon B. Mol Cells Research Article Hepatitis C virus (HCV) propagation is highly dependent on cellular proteins. To identify the host factors involved in HCV propagation, we previously performed protein microarray assays and identified the LIM and SH3 domain protein 1 (LASP-1) as an HCV NS5A-interacting partner. LASP-1 plays an important role in the regulation of cell proliferation, migration, and protein-protein interactions. Alteration of LASP-1 expression has been implicated in hepatocellular carcinoma. However, the functional involvement of LASP1 in HCV propagation and HCV-induced pathogenesis has not been elucidated. Here, we first verified the protein interaction of NS5A and LASP-1 by both in vitro pulldown and coimmunoprecipitation assays. We further showed that NS5A and LASP-1 were colocalized in the cytoplasm of HCV infected cells. NS5A interacted with LASP-1 through the proline motif in domain I of NS5A and the tryptophan residue in the SH3 domain of LASP-1. Knockdown of LASP-1 increased HCV replication in both HCV-infected cells and HCV subgenomic replicon cells. LASP-1 negatively regulated viral propagation and thereby overexpression of LASP-1 decreased HCV replication. Moreover, HCV propagation was decreased by wild-type LASP-1 but not by an NS5A binding-defective mutant of LASP-1. We further demonstrated that LASP-1 was involved in the replication stage of the HCV life cycle. Importantly, LASP-1 expression levels were increased in persistently infected cells with HCV. These data suggest that HCV modulates LASP-1 via NS5A in order to regulate virion levels and maintain a persistent infection. Korean Society for Molecular and Cellular Biology 2020-05-31 2020-04-28 /pmc/articles/PMC7264479/ /pubmed/32344996 http://dx.doi.org/10.14348/molcells.2020.0018 Text en © The Korean Society for Molecular and Cellular Biology. All rights reserved. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
spellingShingle Research Article
Choi, Jae-Woong
Kim, Jong-Wook
Nguyen, Lap P.
Nguyen, Huu C.
Park, Eun-Mee
Choi, Dong Hwa
Han, Kang Min
Kang, Sang Min
Tark, Dongseob
Lim, Yun-Sook
Hwang, Soon B.
Nonstructural NS5A Protein Regulates LIM and SH3 Domain Protein 1 to Promote Hepatitis C Virus Propagation
title Nonstructural NS5A Protein Regulates LIM and SH3 Domain Protein 1 to Promote Hepatitis C Virus Propagation
title_full Nonstructural NS5A Protein Regulates LIM and SH3 Domain Protein 1 to Promote Hepatitis C Virus Propagation
title_fullStr Nonstructural NS5A Protein Regulates LIM and SH3 Domain Protein 1 to Promote Hepatitis C Virus Propagation
title_full_unstemmed Nonstructural NS5A Protein Regulates LIM and SH3 Domain Protein 1 to Promote Hepatitis C Virus Propagation
title_short Nonstructural NS5A Protein Regulates LIM and SH3 Domain Protein 1 to Promote Hepatitis C Virus Propagation
title_sort nonstructural ns5a protein regulates lim and sh3 domain protein 1 to promote hepatitis c virus propagation
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7264479/
https://www.ncbi.nlm.nih.gov/pubmed/32344996
http://dx.doi.org/10.14348/molcells.2020.0018
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