| Vesicular Stomatitis Virus (VSV) is a member of the Rhabdoviridae family that serves as the prototype for highly pathogenic negative-sense non-segmented RNA viruses such as Ebola and rabies viruses and for segmented RNA viruses like influenza. VSV has also proven to be an effective vaccine vector and oncolytic therapy agent. For these reasons, the interaction between VSV and host cells in the context of infection and innate immunity was of great interest. Previous work performed by the laboratory of Douglas Lyles determined that a class of RNA-binding proteins, heterogeneous nuclear ribonucleoproteins (hnRNPs), relocalized from the nucleus to the cytoplasm during infection with VSV. We performed co-immunoprecipitation and RNA-seq experiments to identify and quantify the RNAs that interacted with hnRNP A1, hnRNP C1/C2, and hnRNP K during normal cellular function and during VSV infection. Furthermore, the secondary structure of VSV mRNAs was determined using an experimental structure prediction methodology, Selective 2 Hydroxyl Acylation analyzed by Primer Extension (SHAPE). Our findings detail a preferential interaction between hnRNP C1/C2 and VSV glycoprotein mRNA, and between hnRNP A1 and VSV phosphoprotein mRNA. Additionally, a number of RNA-RNA and RNA-protein binding motifs were identified in VSV nucleoprotein, phosphoprotein and glycoprotein mRNA. |
