Impacts of p97 on Proteome Changes in Human Cells during Coronaviral Replication.

Kai-Wen Cheng,Nallely M Ruiz-Lopez,Nadia Houerbi,Shan Li,Tsui-Fen Chou,Feng Wang

doi:10.3390/cells10112953

Abstract

Human coronavirus (HCoV) similar to other viruses rely on host cell machinery for both replication and to spread. The p97/VCP ATPase is associated with diverse pathways that may favor HCoV replication. In this study, we assessed the role of p97 and associated host responses in human lung cell line H1299 after HCoV-229E or HCoV-OC43 infection. Inhibition of p97 function by small molecule inhibitors shows antiviral activity, particularly at early stages of the virus life cycle, during virus uncoating and viral RNA replication. Importantly, p97 activity inhibition protects human cells against HCoV-induced cytopathic effects. The p97 knockdown also inhibits viral production in infected cells. Unbiased quantitative proteomics analyses reveal that HCoV-OC43 infection resulted in proteome changes enriched in cellular senescence and DNA repair during virus replication. Further analysis of protein changes between infected cells with control and p97 shRNA identifies cell cycle pathways for both HCoV-229E and HCoV-OC43 infection. Together, our data indicate a role for the essential host protein p97 in supporting HCoV replication, suggesting that p97 is a therapeutic target to treat HCoV infection.

Highlights

To firstly identify cell lines that can be infected by both Human coronavirus (HCoV)-229E and HCoV-OC43 viruses we evaluated four cell lines including two lung cancer cell lines, A549 and H1299, as well as the two virus propagating cell lines, MRC-5 and HCT-8
We investigated the effects of HCoV infection on cells and tested whether HCoV229E, HCoV-OC43, and SARS-CoV-2 induce similar changes in cellular processes in infected cells
We find that p97 plays an important role in the life cycle of HCoV

Summary

Introduction

Coronaviruses (CoVs) are a family of enveloped positive-sense single-stranded RNA viruses, that are linked to respiratory and enteric disease [1]. The CoVs are classified under the subfamily Coronovirinae within the family Coronaviridae and the order Nidovirales. Seven human CoVs (HCoVs) have been identified. The HCoV-OC43 and HCoV-HKU1 belong to lineage A, and the SARS-CoV and SARS-CoV-2 belong to lineage B while MERS-CoV belongs to lineage C [2]. Unlike the four common HCoVs (229E, OC43, NL63, and HKU1), which generally cause mild to moderate upper-respiratory tract infections similar to the common cold, SARS-CoV, SARS-CoV-2, and MERS-CoV cause severe, acute respiratory pathologies [3]. The SARS-CoV-2 is highly transmissible and pathogenic and causes coronavirus disease 2019 (COVID-19) [3].

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