Three amino acid substitutions in the NS1 protein change the virus replication of H5N1 influenza virus in human cells

Jing Li; Kun Zhang; Quanjiao Chen; Xiaoshuang Zhang; Yeping Sun; Yuhai Bi; Shuang Zhang; Jinyan Gu; Jiarong Li; Di Liu; Wenjun Liu; Jiyong Zhou

doi:10.1016/j.virol.2018.04.004

Three amino acid substitutions in the NS1 protein change the virus replication of H5N1 influenza virus in human cells

Virology. 2018 Jun:519:64-73. doi: 10.1016/j.virol.2018.04.004. Epub 2018 Apr 17.

Authors

Jing Li¹, Kun Zhang², Quanjiao Chen³, Xiaoshuang Zhang³, Yeping Sun⁴, Yuhai Bi⁴, Shuang Zhang⁴, Jinyan Gu⁵, Jiarong Li⁵, Di Liu⁶, Wenjun Liu⁷, Jiyong Zhou⁸

Affiliations

¹ MOE Joint International Research Laboratory of Animal Immunology, Nanjing Agricultural University, Nanjing, China; CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
² Philips Institute for Oral Health Research, School of Dentistry, Virginia Commonwealth University, Richmond, VA, USA.
³ CAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Hubei, China.
⁴ CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
⁵ MOE Joint International Research Laboratory of Animal Immunology, Nanjing Agricultural University, Nanjing, China.
⁶ CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China; CAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Hubei, China.
⁷ CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China. Electronic address: liuwj@im.ac.cn.
⁸ MOE Joint International Research Laboratory of Animal Immunology, Nanjing Agricultural University, Nanjing, China; Key Laboratory of Animal Virology, Ministry of Agriculture, Zhejiang University, Hangzhou, China; Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University, Hangzhou, China. Electronic address: jyzhou@zju.edu.cn.

PMID: 29677653
DOI: 10.1016/j.virol.2018.04.004

Abstract

Influenza A viruses have sophisticated strategies to promote their own replication. Here, we found that three H5N1 influenza viruses display different replication patterns in human A549 and macrophage cells. The HN01 virus displayed poor replication compared to HN021 and JS01. In addition, the HN01 virus was unable to counteract the interferon response and block general gene expression. This capability was restored by three amino acid substitutions on the NS1 protein: K55E, K66E, and C133F, resulting in recovered binding to CPSF30 and decreased interferon response activity. Furthermore, a recombinant HN01 virus expressing either NS1-C133F or the triple mutation replicate with higher titers in human A549 cells and macrophages compared to the parent virus. These three amino acid mutations reveal a new pathway to alter H5N1 virus replication.

Keywords: H5N1 influenza virus; IFN response; NS1 protein; Virus replication.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

A549 Cells
Amino Acid Substitution*
Animals
Chemokines / biosynthesis
Chemokines / genetics
Cytokines / biosynthesis
Cytokines / genetics
Dogs
Host-Pathogen Interactions
Humans
Immunity, Innate
Influenza A Virus, H1N1 Subtype / genetics
Influenza A Virus, H5N1 Subtype / genetics*
Influenza A Virus, H5N1 Subtype / physiology*
Interferons
Macrophages / virology*
Madin Darby Canine Kidney Cells
Viral Nonstructural Proteins / chemistry
Viral Nonstructural Proteins / genetics*
Viral Nonstructural Proteins / metabolism
Virulence
Virus Replication*

Substances

Chemokines
Cytokines
INS1 protein, influenza virus
Viral Nonstructural Proteins
Interferons