The chemokine receptor homologue encoded by US27 of human cytomegalovirus is heavily glycosylated and is present in infected human foreskin fibroblasts and enveloped virus particles

Barry J. Margulies, Wade Gibson

Research output: Contribution to journalArticle

Abstract

Human cytomegalovirus (HCMV), a member of the β-herpesvirus family, encodes four homologues of cellular G protein-coupled receptors (GPCRs). One of these, the protein product of HCMV open reading frame (ORF) UL33, has been identified in HCMV-infected cells and virus particles and shown to be heat-aggregatable and N-glycosylated. Another, the product of ORF US28, has been functionally characterized as a β-chemokine receptor. Here we report the use of RT-PCR, coupled in vitro transcription-translation, immunoprecipitation, and Western immunoassays to (i) show that RNA from the open reading frame US27 appears predominantly during the late phase of replication; (ii) identify the protein encoded by HCMV US27 in infected cells and enveloped virus particles; (iii) demonstrate that the US27-encoded protein is heterogeneously N-glycosylated and resolves as two species following treatment with peptide N-glycosidase F; and (iv) show that both the recombinant and deglycoylated infected cell US27 protein aggregate when heated in the presence of SDS prior to electrophoresis in polyacrylamide gels, a property which is abrogated with the addition of urea to sample buffer.

Original languageEnglish (US)
Pages (from-to)57-71
Number of pages15
JournalVirus Research
Volume123
Issue number1
DOIs
StatePublished - Jan 1 2007

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Keywords

  • Chemokine receptor
  • Cytomegalovirus
  • Signal transduction
  • UL33
  • UL78
  • US28
  • Viral homologue

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases
  • Cancer Research

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