title: An unbiased proteomics approach to identify human cytomegalovirus RNA-associated proteins.
abstract: Post-transcriptional events regulate herpesvirus gene expression, yet few herpesvirus RNA-binding proteins have been identified. We used an unbiased approach coupling oligo(dT) affinity capture with proteomics to identify viral RNA-associated proteins during infection. Using this approach, we identified and confirmed changes in the abundance or activity of two host RNA-associated proteins, DHX9 and DDX3, in cells infected with human cytomegalovirus (HCMV). We also identified and confirmed previously unreported activities for the HCMV US22 and pp71 proteins as RNA-associated viral proteins and confirmed that a known viral RNA-binding protein, pTRS1, associates with RNA in infected cells. Further, we found that HCMV pp71 co-sedimented with polysomes, associated with host and viral RNAs, and stimulated the overall rate of protein synthesis. These results demonstrate that oligo(dT) affinity capture coupled with proteomics provides a rapid and straightforward means to identify RNA-associated viral proteins during infection that may participate in the post-transcriptional control of gene expression.
authors.affiliation: Department of Microbiology & Immunology, Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC, United States.
authors.forename: Erik M
authors.initials: EM
authors.lastname: Lenarcic
authors.affiliation: Department of Microbiology & Immunology, Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC, United States.
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Ziehr
authors.affiliation: Department of Microbiology & Immunology, Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC, United States. Electronic address: nmoorman@med.unc.edu.
authors.forename: Nathaniel J
authors.initials: NJ
authors.lastname: Moorman
mesh_terms: D003587:Cytomegalovirus / Q000235:genetics / Q000378:metabolism; D003586:Cytomegalovirus Infections / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D006801:Humans; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D040901:Proteomics; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D012367:RNA, Viral; D016601:RNA-Binding Proteins
keywords: Herpesvirus; Human cytomegalovirus (HCMV); Messenger RNA (mRNA); Post-transcriptional regulation; Proteomics; RNA-binding; Translation