An unbiased proteomics approach to identify human cytomegalovirus RNA-associated proteins

Virology
Erik M LenarcicNathaniel J Moorman

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.

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Citations

Apr 28, 2016·Proceedings of the National Academy of Sciences of the United States of America·Annalaura BraiMaurizio Botta
Feb 27, 2017·Molecular & Cellular Proteomics : MCP·Kyle C ArendNathaniel J Moorman
Jan 29, 2016·Journal of Virology·Benjamin ZiehrNathaniel J Moorman
Apr 2, 2016·Oncotarget·Teresa Lee, Jerry Pelletier
Mar 17, 2018·Virus Genes·Ana Carla Peixoto GuissoniDivina das Dores de Paula Cardoso
Apr 1, 2020·Frontiers in Cellular and Infection Microbiology·Robert F Kalejta, Emily R Albright
Dec 7, 2018·Cell Death and Differentiation·Xiangya DingChun Lu
Dec 11, 2020·Antiviral Research·Paul T WinnardVenu Raman
Sep 21, 2019·Virology·Heather A VincentNathaniel J Moorman
Feb 16, 2021·Frontiers in Cellular and Infection Microbiology·Kai-Min LinMichael P Weekes
Nov 3, 2020·Virus Research·Fan Guo, Li Xing
May 8, 2021·Journal of Proteome Research·Flora CozzolinoMaria Monti
Jul 3, 2021·International Journal of Molecular Sciences·Olga SergeevaTimofei Zatsepin

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