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PMID: 8918538 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Identification of the human cytomegalovirus G protein-coupled receptor homologue encoded by UL33 in infected cells and enveloped virus particles.

Virology ·Vol. 225 ·No. 1 ·1996-11-01 ·Pages 111-25

Margulies BJ, Browne H, Gibson W

Abstract

Human cytomegalovirus (HCMV), strain AD169, contains four genes (US27, US28, UL33, and UL78) that encode putative homologues of cellular G protein-coupled receptors (GCRs). GCRs transduce extracellular signals to alter intracellular processes, and there is evidence that HCMV may elicit such changes at early times following infection. The US27, US28, and UL33 genes are transcribed during infection, and the US28 gene product has been found to be a functional receptor for the beta-chemokine class of immune modulators. The US27, UL33, and UL78 gene products have not been described and we have concentrated on identifying the UL33 protein because it is the most highly conserved of the GCR homologues among the human beta and gamma herpesviruses. We report here cloning UL33 into a recombinant baculovirus (rBV) and expressing it in insect cells; constructing a mutant HCMV with a disrupted UL33 gene; and identifying the UL33 protein in HCMV-infected cells and virus particles. Our results demonstrate that the UL33 protein (i) is expressed as a approximately 36-kDa, heat-aggregatable protein in rBV-infected cells, (ii) is modified heterogeneously by asparagine-linked glycosylation and expressed as a > or = 58-kDa glycoprotein that is present in the region of the cytoplasmic inclusions in HCMV-infected fibroblasts, (iii) is present in virions and two other enveloped virus particles, and (iv) is not essential for growth of HCMV in human foreskin fibroblast cultures.

MeSH Terms
Amidohydrolases Amino Acid Sequence Animals Asparagine/metabolism Cell Line Cloning, Molecular Cytomegalovirus/chemistry,genetics,growth & development Fibroblasts/virology GTP-Binding Proteins Glycosylation Humans Inclusion Bodies, Viral/virology Molecular Sequence Data Molecular Weight Nucleopolyhedroviruses/genetics Peptide Fragments/analysis Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase RNA Splicing Receptors, Cell Surface/genetics Recombinant Proteins Sequence Homology, Amino Acid Spodoptera Viral Proteins/analysis,chemistry,genetics,physiology Virion/chemistry
Chemicals
Peptide Fragments Receptors, Cell Surface Recombinant Proteins Viral Proteins Asparagine Amidohydrolases Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase GTP-Binding Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Margulies B J
Retrovirus Biology Laboratory, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Browne H
Gibson W
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Article Info
Journal
Virology
Abbr.
Virology
ISSN
0042-6822
DOI
10.1006/viro.1996.0579
Published
1996-11-01
Pages
111-25
Language
English
Region
United States
NLM ID
0110674
PMCID
PMC6953178
Subset
IM
Grants
NIAID NIH HHS · AI22711 · United States
NIAID NIH HHS · R01 AI013718-23 · United States
NIAID NIH HHS · R01 AI013718 · United States
NIAID NIH HHS · AI13718 · United States
NIGMS NIH HHS · GM07445 · United States
NIAID NIH HHS · R01 AI013718-24 · United States
Wellcome Trust · United Kingdom
NIAID NIH HHS · R01 AI013718-25 · United States
External Links
PubMed source DOI: 10.1006/viro.1996.0579 PMC Full Text View journal details
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