Human cytomegalovirus TRS1 protein associates with the 7-methylguanosine mRNA cap and facilitates translation.

TitleHuman cytomegalovirus TRS1 protein associates with the 7-methylguanosine mRNA cap and facilitates translation.
Publication TypeJournal Article
Year of Publication2015
AuthorsZiehr, B, Lenarcic, E, Vincent, HA, Cecil, C, Garcia, B, Shenk, T, Moorman, NJ
JournalProteomics
Volume15
Issue12
Pagination1983-94
Date Published2015 Jun 01
ISSN1615-9861
KeywordsBlotting, Western, Cells, Cultured, Cytomegalovirus, Cytomegalovirus Infections, Fibroblasts, Gene Expression Regulation, Viral, Guanosine, HeLa Cells, Humans, Immunoprecipitation, Protein Biosynthesis, Ribosomes, RNA Cap-Binding Proteins, RNA, Messenger, RNA, Viral, Viral Proteins, Virus Replication
Abstract

Viruses rely on the host translation machinery for the synthesis of viral proteins. Human cells have evolved sensors that recognize viral RNAs and inhibit mRNA translation in order to limit virus replication. Understanding how viruses manipulate the host translation machinery to gain access to ribosomes and disable the antiviral response is therefore a critical aspect of the host/pathogen interface. In this study, we used a proteomics approach to identify human cytomegalovirus (HCMV) proteins that might contribute to viral mRNA translation. The HCMV TRS1 protein (pTRS1) associated with the 7-methylguanosine mRNA cap, increased the total level of protein synthesis, and colocalized with mRNAs undergoing translation initiation during infection. pTRS1 stimulated translation of a nonviral reporter gene and increased the translation of a reporter containing an HCMV 5' untranslated region (5'UTR) to a greater extent. The preferential effect of pTRS1 on translation of an mRNA containing a viral 5'UTR required the pTRS1 RNA and double-stranded RNA-dependent protein kinase (PKR)-binding domains, and was likely the result of PKR inhibition. However, pTRS1 also stimulated the total level of protein synthesis and translation directed by an HCMV 5'UTR in cells lacking PKR. Thus our results demonstrate that pTRS1 stimulates translation through both PKR-dependent and PKR-independent mechanisms.

DOI10.1002/pmic.201400616
Alternate JournalProteomics
PubMed ID25894605
PubMed Central IDPMC4492309
Grant ListT32 AI007419 / AI / NIAID NIH HHS / United States
AI103311-02S1 / AI / NIAID NIH HHS / United States
P30 CA016086 / CA / NCI NIH HHS / United States
R56 AI112951 / AI / NIAID NIH HHS / United States
AI112951 / AI / NIAID NIH HHS / United States
AI103311 / AI / NIAID NIH HHS / United States
R01 AI112951 / AI / NIAID NIH HHS / United States
R01 AI103311 / AI / NIAID NIH HHS / United States