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: Erik M Lenarcic, Benjamin J Ziehr, Nathaniel J Moorman
• Journal: Virology (Virology) Vol. 481 Pg. 13-23 (Jul 2015) ISSN: 1096-0341 [Electronic] United States
• PMID: 25765003 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Published by Elsevier Inc.

Chemical References

• RNA, Viral
• RNA-Binding Proteins

Topics

• Cytomegalovirus (genetics, metabolism)
• Cytomegalovirus Infections (genetics, metabolism, virology)
• Humans
• Molecular Sequence Data
• Protein Biosynthesis
• Proteomics
• RNA, Viral (genetics, metabolism)
• RNA-Binding Proteins (chemistry, genetics, metabolism)