Abstract

Although orthopoxviruses (OPXV) are known to encode a majority of the genes required for replication in host cells, genome-wide genetic screens have revealed that several host pathways are indispensable for OPXV infection. Through a haploid genetic screen, we previously identified several host genes required for monkeypox virus (MPXV) infection, including the individual genes that form the conserved oligomeric Golgi (COG) complex. The COG complex is an eight-protein (COG1–COG8) vesicle tethering complex important for regulating membrane trafficking, glycosylation enzymes, and maintaining Golgi structure. In this study, we investigated the role of the COG complex in OPXV infection using cell lines with individual COG gene knockout (KO) mutations. COG KO cells infected with MPXV and vaccinia virus (VACV) produced small plaques and a lower virus yield compared to wild type (WT) cells. In cells where the KO phenotype was reversed using a rescue plasmid, the size of virus plaques increased demonstrating a direct link between the decrease in viral spread and the KO of COG genes. KO cells infected with VACV displayed lower levels of viral fusion and entry compared to WT suggesting that the COG complex is important for early events in OPXV infection. Additionally, fewer actin tails were observed in VACV-infected KO cells compared to WT. Since COG complex proteins are required for cellular trafficking of glycosylated membrane proteins, the disruption of this process due to lack of individual COG complex proteins may potentially impair the virus-cell interactions required for viral entry and egress. These data validate that the COG complex previously identified in our genetic screens plays a role in OPXV infection.

Highlights

  • The orthopoxvirus (OPXV) genus includes several viruses that can infect humans, such as variola virus, monkeypox virus and vaccinia virus

  • We showed that the absence of conserved oligomeric Golgi (COG) proteins caused a decrease in the number of actin tails in infected cells, and decreased viral spread, as evidenced by smaller vaccinia virus (VACV) and monkeypox virus (MPXV) plaques produced in COG KO cells compared to wildtype (WT) cells

  • Since genes from both lobes were significantly enriched in the previous haploid screen, we further investigated the role of all COG complex genes in OPXV infection

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Summary

Introduction

The orthopoxvirus (OPXV) genus includes several viruses that can infect humans, such as variola virus (the causative agent of eradicated smallpox), monkeypox virus (an emerging zoonotic infectious pathogen) and vaccinia virus (the prototypic poxvirus member extensively used in research studies and vaccines). Infection begins with viral entry, a multistep process that includes virus attachment to the host cell, fusion of viral and host membranes, and core entry into the cytoplasm. These early events in infection can vary depending on the infectious form of the virus, the virus strain, and the host target cell type, among other factors [1,2]. The initial attachment of virus to cells involves viral proteins that target host cell glycosaminoglyans (GAGs) or laminins [3,4,5]

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