Vault RNAs aid RNA virus infection by facilitating cytoplasmic localization of hnRNP C and ELAVL1

Vault RNAs (vtRNAs) are a family of four small non-coding RNAs that regulate multiple cellular pathways in human cells. VtRNA expression is increased upon infection with certain RNA and DNA viruses, suggesting that they either promote or restrict viral replication. However, their precise molecular function remains unclear. We show that replication of picornaviruses, alphaviruses, and beta-coronaviruses broadly enhances vtRNA expression. Genetic loss of vtRNAs inhibits replication of Sindbis virus (SINV) and encephalomyocarditis virus (EMCV), independent of the antiviral type I interferon response. A proteomic screen demonstrates that vtRNAs associate with RNA-binding proteins ELAVL1 and hnRNP C in uninfected and infected cells. During infection, vtRNAs facilitate cytoplasmic localization of ELAVL1 and hnRNP C, where these proteins associate with viral RNA and support SINV replication. Together, our findings reveal a proviral function of vtRNAs during SINV and EMCV infection, which presents therapeutic opportunities to fight infectious diseases.