Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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Nonstructural NS5A Protein Regulates LIM and SH3 Domain Protein 1 to Promote Hepatitis C Virus Propagation

  • Choi, Jae-Woong (Laboratory of RNA Viral Diseases, Korea Zoonosis Research Institute, Jeonbuk National University) ;
  • Kim, Jong-Wook (Ilsong Institute of Life Science, Hallym University) ;
  • Nguyen, Lap P. (Laboratory of RNA Viral Diseases, Korea Zoonosis Research Institute, Jeonbuk National University) ;
  • Nguyen, Huu C. (Laboratory of RNA Viral Diseases, Korea Zoonosis Research Institute, Jeonbuk National University) ;
  • Park, Eun-Mee (Center for Immunology and Pathology, National Institute of Health, Korea Center for Disease Control & Prevention) ;
  • Choi, Dong Hwa (Biocenter, Gyeonggido Business & Science Accelerator) ;
  • Han, Kang Min (Department of Pathology, Dongguk University Ilsan Hospital) ;
  • Kang, Sang Min (Laboratory for Infectious Disease Prevention, Korea Zoonosis Research Institute, Jeonbuk National University) ;
  • Tark, Dongseob (Laboratory for Infectious Disease Prevention, Korea Zoonosis Research Institute, Jeonbuk National University) ;
  • Lim, Yun-Sook (Laboratory of RNA Viral Diseases, Korea Zoonosis Research Institute, Jeonbuk National University) ;
  • Hwang, Soon B. (Laboratory of RNA Viral Diseases, Korea Zoonosis Research Institute, Jeonbuk National University)
  • Received : 2020.01.14
  • Accepted : 2020.03.20
  • Published : 2020.05.31
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Abstract

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 LASP-1 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 LASP1 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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References

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