Global analysis of polysome-associated mRNA in vesicular stomatitis virus infected cells

Abstract

Infection of mammalian cells with vesicular stomatitis virus (VSV) results in the inhibition of cellular translation while viral translation proceeds efficiently. VSV RNA synthesis occurs entirely within the cytoplasm, where during transcription the viral polymerase produces 5 mRNAs that are structurally indistinct to cellular mRNAs with respect to their 5′ cap-structure and 3′-polyadenylate tail. Using the global approach of massively parallel sequencing of total cytoplasmic, monosome- and polysome-associated mRNA, we interrogate the impact of VSV infection of HeLa cells on translation. Analysis of sequence reads in the different fractions shows >60% of total cytoplasmic and polysome-associated reads map to the 5 viral genes by 6 hours post-infection, a time point at which robust host cell translational shut-off is observed. Consistent with an overwhelming abundance of viral mRNA in the polysome fraction, the reads mapping to cellular genes were reduced. The cellular mRNAs that remain most polysome-associated following infection had longer half-lives, were typically larger, and were more AU rich, features that are shared with the viral mRNAs. Several of those mRNAs encode proteins known to positively affect viral replication, and using chemical inhibition and siRNA depletion we confirm that the host chaperone heat shock protein 90 (hsp90) and eukaryotic translation initiation factor 3A (eIF3A)—encoded by 2 such mRNAs—support viral replication. Correspondingly, regulated in development and DNA damage 1 (Redd1) encoded by a host mRNA with reduced polysome association inhibits viral infection. These data underscore the importance of viral mRNA abundance in the shut-off of host translation in VSV infected cells and link the differential translatability of some cellular mRNAs with pro- or antiviral function.

The proximal proteome of 17 SARS-CoV-2 proteins links to disrupted antiviral signaling and host translation

“…This element exists on all genomic and subgenomic viral RNAs (71). Whether other SARS-CoV-2 factors are necessary to overcome NSP1 translational inhibition or if, during viral replication, the large number of viral transcripts simply outcompetes host transcripts, as seen in vesicular stomatitis virus (72), remains to be determined. A recent study(73) from autopsies of COVID patients characterized whole proteome changes in multiple organs.…”

Effect of the Viral Hemorrhagic Septicemia Virus Nonvirion Protein on Translation via PERK-eIF2α Pathway

“…The impact of VSV on host translation during infection has been extensively studied. Studies proposed that VSV alters the eIF4F translation initiation complex to inhibit host protein synthesis and that the abundance of viral mRNA during VSV infection contributes to the host cell shutoff [47][48][49]. VHSV IVb may be impacting host translational shutoff in a similar manner to VSV and further studies to better identify the exact mechanism are required.…”

Components and Architecture of the Rhabdovirus Ribonucleoprotein Complex

“…Furthermore, an interaction of the first 10 N-terminal amino acids with dynamin is essential for virus assembly [25] and the N-terminus of the M-protein is also responsible for the interaction with the N-protein [26] and lipid membranes [27,28]. Apart from its roles in virus assembly, the VSV M-protein suppresses host mRNA transcription and translation by interfering with the export of mRNAs from the nucleus [10,[29][30][31] and the phosphorylation of transcription factors [32][33][34][35]. Additionally, the VSV M-protein has been shown to affect immunoproteasome formation via interaction with LMP2 [36].…”