RNA‐Protein Interactions Involved in Persistence of Cytomegalovirus

Abstract

Human Cytomegalovirus (HCMV) is a ubiquitous herpes virus and usually asymptomatic among healthy individuals. Congenital HCMV infections are the leading cause of virally induced birth defects and HCMV causes severe disease in immunocompromised patients. Noncoding RNA is a class of RNA molecules that have a wide range of structures and functions which can exhibit cell‐specified specialization and expression. HCMV encodes many ncRNAs that range in size from small (miRNAs) to large stable introns. HCMV encodes several long noncoding RNAs (lncRNAs) of various function. Previous work has shown that HCMV encodes a lncRNA called RNA5.0. The ortholog of RNA5.0 in the MCVM, RNA7.2, has shown to play a role in virus persistence within host salivary glands. However, the molecular interactions/mechanism of this process remains unclear. In order to begin characterizing the molecular function of RNA5.0, we will use the RaPID system to screen for RNA‐protein interactions. The RaPID system recruits a highly active biotin ligase to tag proteins bound on the RNA. Previous work demonstrated that the 3′ end of RNA5.0 is related to the processing and stability of the RNA. There is a hairpin loop that is absolutely required for RNA stability. Several different length fragments spanning the hairpin loop have been cloned into the RaPID system. These constructs will be used to purify RNA binding proteins. Mass spectrometry analysis will be used to identify specific HCMV RNA binding proteins. The identified binding proteins might point to specific pathways involved in viral persistence that could be targeted for potential treatment for CMV.