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 language | English (US) |
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Pages (from-to) | 57-71 |
Number of pages | 15 |
Journal | Virus Research |
Volume | 123 |
Issue number | 1 |
DOIs | |
State | Published - Jan 2007 |
Keywords
- Chemokine receptor
- Cytomegalovirus
- Signal transduction
- UL33
- UL78
- US28
- Viral homologue
ASJC Scopus subject areas
- Virology
- Infectious Diseases
- Cancer Research