Viral-Host Interactome Analysis Reveals Chicken STAU2 Interacts With Non-structural Protein 1 and Promotes the Replication of H5N1 Avian Influenza Virus.

Qiao Wang,Maiqing Zheng,Qinghe Li,Guiping Zhao,Jie Wen,Qi Zhang

doi:10.3389/fimmu.2021.590679

Abstract

As a highly pathogenic influenza virus, H5N1 avian influenza virus (AIV) poses a great threat to poultry production and public health. H5N1 AIV has a small genome and, therefore, relies heavily on its host cellular machinery to replicate. To develop a comprehensive understanding of how H5N1 AIV rewires host cellular machinery during the course of infection, it is crucial to identify which host proteins and complexes come into physical contact with the viral proteins. Here, we utilized affinity purification mass spectrometry (AP-MS) to systematically determine the physical interactions of 11 H5N1 AIV proteins with host proteins in chicken DF1 cells. We identified with high confidence 1,043 H5N1 AIV–chicken interactions involving 621 individual chicken proteins and uncovered a number of host proteins and complexes that were targeted by the viral proteins. Specifically, we revealed that chicken Staufen double-stranded RNA-binding protein 2 interacts with AIV non-structural protein 1 (NS1) and promotes the replication of the virus by enhancing the nuclear export of NS1 mRNA. This dataset facilitates a more comprehensive and detailed understanding of how the host machinery is manipulated during the course of H5N1 AIV infection.

Highlights

Influenza A virus (IAV) is a segmented, single-stranded, negative-sense RNA virus that has adapted to infect multiple species
We aimed to systematically and quantitatively identify host proteins associated with H5N1 avian influenza virus (AIV) proteins using affinity purification mass spectrometry (AP-MS)
Affinity purification was carried out using FLAG tag or GFP tag antibodies to immunoprecipitate the host proteins associated with the tagged viral proteins

Summary

Introduction

Influenza A virus (IAV) is a segmented, single-stranded, negative-sense RNA virus that has adapted to infect multiple species. This virus causes annual epidemics and recurring pandemics, which have huge impacts on public health. IAV particles have two viral surface glycoproteins (hemagglutinin, HA; neuraminidase, NA) and one matrix-2 protein (M2). All eight viral RNA (vRNA) segments bind three RNA polymerases (polymerase acid protein, PA; polymerase basic protein 1, PB1; and 2, PB2) and are encapsidated by the nucleoprotein (NP) to form the viral ribonucleoprotein (vRNP) complexes [1,2,3,4,5]. The vRNPs are surrounded by a layer of the matrix protein, M1, which lines the envelope [6]. The host cells detect the viral RNA through pathogen sensors, and the major gene products of the influenza virus mediate the viral life cycle and modulate cellular processes [7]

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