Double-stranded RNA binding by human cytomegalovirus pTRS1

Morgan Hakki, Adam P. Geballe

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

The human cytomegalovirus (HCMV) TRS1 and IRS1 genes rescue replication of vaccinia virus (VV) that has a deletion of the double-stranded RNA binding protein gene E3L (VVΔE3L). Like E3L, these HCMV genes block the activation of key interferon-induced, double-stranded RNA (dsRNA)-activated antiviral pathways. We investigated the hypothesis that the products of these HCMV genes act by binding to dsRNA. pTRS1 expressed by cell-free translation or by infection of mammalian cells with HCMV or recombinant VV bound to dsRNA. Competition experiments revealed that pTRS1 preferentially bound to dsRNA compared to double-stranded DNA or single-stranded RNA. 5′- and 3′-end deletion analyses mapped the TRS1 dsRNA-binding domain to amino acids 74 through 248, a region of identity to pIRS1 that contains no homology to known dsRNA-binding proteins. Deletion of the majority of this region (Δ86-246) completely abrogated dsRNA binding. To determine the role of the dsRNA-binding domain in the rescue of VVΔE3L replication, wild-type or deletion mutants of TRS1 were transfected into HeLa cells, which were then infected with VVΔE3L. While full-length TRS1 rescued VVΔE3L replication, deletion mutants affecting a carboxy-terminal region of TRS1 that is not required for dsRNA binding failed to rescue VVΔE3L. Analyses of stable cell lines revealed that the carboxy-terminal domain is necessary to prevent the shutoff of protein synthesis and the phosphorylation of eIF2α after VVΔE3L infection. Thus, pTRS1 contains an unconventional dsRNA-binding domain at its amino terminus, but a second function involving the car boxy terminus is also required for countering host cell antiviral responses.

Original languageEnglish (US)
Pages (from-to)7311-7318
Number of pages8
JournalJournal of Virology
Volume79
Issue number12
DOIs
StatePublished - Jun 2005
Externally publishedYes

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Human herpesvirus 5
Double-Stranded RNA
double-stranded RNA
Cytomegalovirus
RNA-Binding Proteins
Vaccinia virus
Antiviral Agents
Genes
RNA-binding proteins
Infection
HeLa Cells
Interferons
Transcriptional Activation
cells
Phosphorylation
RNA
mutants
Amino Acids
gene activation
genes

ASJC Scopus subject areas

  • Immunology

Cite this

Double-stranded RNA binding by human cytomegalovirus pTRS1. / Hakki, Morgan; Geballe, Adam P.

In: Journal of Virology, Vol. 79, No. 12, 06.2005, p. 7311-7318.

Research output: Contribution to journalArticle

Hakki, Morgan ; Geballe, Adam P. / Double-stranded RNA binding by human cytomegalovirus pTRS1. In: Journal of Virology. 2005 ; Vol. 79, No. 12. pp. 7311-7318.
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