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Journal of Virology logoLink to Journal of Virology
. 1997 Feb;71(2):1521–1529. doi: 10.1128/jvi.71.2.1521-1529.1997

Identification and characterization of a G protein-coupled receptor homolog encoded by murine cytomegalovirus.

N J Davis-Poynter 1, D M Lynch 1, H Vally 1, G R Shellam 1, W D Rawlinson 1, B G Barrell 1, H E Farrell 1
PMCID: PMC191209  PMID: 8995678

Abstract

This report describes the identification of a murine cytomegalovirus (MCMV) G protein-coupled receptor (GCR) homolog. This open reading frame (M33) is most closely related to, and collinear with, human cytomegalovirus UL33, and homologs are also present in human herpesvirus 6 and 7 (U12 for both viruses). Conserved counterparts in the sequenced alpha- or gammaherpesviruses have not been identified to date, suggesting that these genes encode proteins which are important for the biological characteristics of betaherpesviruses. We have detected transcripts for both UL33 and M33 as early as 3 or 4 h postinfection, and these reappear at late times. In addition, we have identified N-terminal splicing for both the UL33 and M33 RNA transcripts. For both open reading frames, splicing results in the introduction of amino acids which are highly conserved among known GCRs. To characterise the function of the M33 in the natural host, two independent MCMV recombinant viruses were prepared, each of which possesses an M33 open reading frame which has been disrupted with the beta-galactosidase gene. While the recombinant M33 null viruses showed no phenotypic differences in replication from wild-type MCMV in primary mouse embryo fibroblasts in vitro, they showed severely restricted growth in the salivary glands of infected mice. These data suggest that M33 plays an important role in vivo, in particular in the dissemination to or replication in the salivary gland, and provide the first evidence for the function of a viral GCR homolog in vivo.

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Selected References

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  1. Ahuja S. K., Murphy P. M. Molecular piracy of mammalian interleukin-8 receptor type B by herpesvirus saimiri. J Biol Chem. 1993 Oct 5;268(28):20691–20694. [PubMed] [Google Scholar]
  2. Allan J. E., Shellam G. R. Genetic control of murine cytomegalovirus infection: virus titres in resistant and susceptible strains of mice. Arch Virol. 1984;81(1-2):139–150. doi: 10.1007/BF01309303. [DOI] [PubMed] [Google Scholar]
  3. Birnstiel M. L., Busslinger M., Strub K. Transcription termination and 3' processing: the end is in site! Cell. 1985 Jun;41(2):349–359. doi: 10.1016/s0092-8674(85)80007-6. [DOI] [PubMed] [Google Scholar]
  4. Brown J. M., Kaneshima H., Mocarski E. S. Dramatic interstrain differences in the replication of human cytomegalovirus in SCID-hu mice. J Infect Dis. 1995 Jun;171(6):1599–1603. doi: 10.1093/infdis/171.6.1599. [DOI] [PubMed] [Google Scholar]
  5. Cao J. X., Gershon P. D., Black D. N. Sequence analysis of HindIII Q2 fragment of capripoxvirus reveals a putative gene encoding a G-protein-coupled chemokine receptor homologue. Virology. 1995 May 10;209(1):207–212. doi: 10.1006/viro.1995.1244. [DOI] [PubMed] [Google Scholar]
  6. Cardin R. D., Abenes G. B., Stoddart C. A., Mocarski E. S. Murine cytomegalovirus IE2, an activator of gene expression, is dispensable for growth and latency in mice. Virology. 1995 May 10;209(1):236–241. doi: 10.1006/viro.1995.1249. [DOI] [PubMed] [Google Scholar]
  7. Cavanaugh V. J., Stenberg R. M., Staley T. L., Virgin H. W., 4th, MacDonald M. R., Paetzold S., Farrell H. E., Rawlinson W. D., Campbell A. E. Murine cytomegalovirus with a deletion of genes spanning HindIII-J and -I displays altered cell and tissue tropism. J Virol. 1996 Mar;70(3):1365–1374. doi: 10.1128/jvi.70.3.1365-1374.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chee M. S., Satchwell S. C., Preddie E., Weston K. M., Barrell B. G. Human cytomegalovirus encodes three G protein-coupled receptor homologues. Nature. 1990 Apr 19;344(6268):774–777. doi: 10.1038/344774a0. [DOI] [PubMed] [Google Scholar]
  9. Chen C., Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. doi: 10.1128/mcb.7.8.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  11. Forrester A., Farrell H., Wilkinson G., Kaye J., Davis-Poynter N., Minson T. Construction and properties of a mutant of herpes simplex virus type 1 with glycoprotein H coding sequences deleted. J Virol. 1992 Jan;66(1):341–348. doi: 10.1128/jvi.66.1.341-348.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gao J. L., Murphy P. M. Human cytomegalovirus open reading frame US28 encodes a functional beta chemokine receptor. J Biol Chem. 1994 Nov 18;269(46):28539–28542. [PubMed] [Google Scholar]
  13. Gayle R. B., 3rd, Sleath P. R., Srinivason S., Birks C. W., Weerawarna K. S., Cerretti D. P., Kozlosky C. J., Nelson N., Vanden Bos T., Beckmann M. P. Importance of the amino terminus of the interleukin-8 receptor in ligand interactions. J Biol Chem. 1993 Apr 5;268(10):7283–7289. [PubMed] [Google Scholar]
  14. Gompels U. A., Nicholas J., Lawrence G., Jones M., Thomson B. J., Martin M. E., Efstathiou S., Craxton M., Macaulay H. A. The DNA sequence of human herpesvirus-6: structure, coding content, and genome evolution. Virology. 1995 May 10;209(1):29–51. doi: 10.1006/viro.1995.1228. [DOI] [PubMed] [Google Scholar]
  15. Henson D., Strano A. J. Mouse cytomegalovirus. Necrosis of infected and morphologically normal submaxillary gland acinar cells during termination of chronic infection. Am J Pathol. 1972 Jul;68(1):183–202. [PMC free article] [PubMed] [Google Scholar]
  16. Hudson J. B. The murine cytomegalovirus as a model for the study of viral pathogenesis and persistent infections. Arch Virol. 1979;62(1):1–29. doi: 10.1007/BF01314900. [DOI] [PubMed] [Google Scholar]
  17. Lagenaur L. A., Manning W. C., Vieira J., Martens C. L., Mocarski E. S. Structure and function of the murine cytomegalovirus sgg1 gene: a determinant of viral growth in salivary gland acinar cells. J Virol. 1994 Dec;68(12):7717–7727. doi: 10.1128/jvi.68.12.7717-7727.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Liu D. X., Cavanagh D., Green P., Inglis S. C. A polycistronic mRNA specified by the coronavirus infectious bronchitis virus. Virology. 1991 Oct;184(2):531–544. doi: 10.1016/0042-6822(91)90423-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Manning W. C., Stoddart C. A., Lagenaur L. A., Abenes G. B., Mocarski E. S. Cytomegalovirus determinant of replication in salivary glands. J Virol. 1992 Jun;66(6):3794–3802. doi: 10.1128/jvi.66.6.3794-3802.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Massung R. F., Jayarama V., Moyer R. W. DNA sequence analysis of conserved and unique regions of swinepox virus: identification of genetic elements supporting phenotypic observations including a novel G protein-coupled receptor homologue. Virology. 1993 Dec;197(2):511–528. doi: 10.1006/viro.1993.1625. [DOI] [PubMed] [Google Scholar]
  21. Murphy P. M. The molecular biology of leukocyte chemoattractant receptors. Annu Rev Immunol. 1994;12:593–633. doi: 10.1146/annurev.iy.12.040194.003113. [DOI] [PubMed] [Google Scholar]
  22. Neote K., DiGregorio D., Mak J. Y., Horuk R., Schall T. J. Molecular cloning, functional expression, and signaling characteristics of a C-C chemokine receptor. Cell. 1993 Feb 12;72(3):415–425. doi: 10.1016/0092-8674(93)90118-a. [DOI] [PubMed] [Google Scholar]
  23. Nicholas J., Cameron K. R., Honess R. W. Herpesvirus saimiri encodes homologues of G protein-coupled receptors and cyclins. Nature. 1992 Jan 23;355(6358):362–365. doi: 10.1038/355362a0. [DOI] [PubMed] [Google Scholar]
  24. Nicholas J. Determination and analysis of the complete nucleotide sequence of human herpesvirus. J Virol. 1996 Sep;70(9):5975–5989. doi: 10.1128/jvi.70.9.5975-5989.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nicholas J., Martin M. E. Nucleotide sequence analysis of a 38.5-kilobase-pair region of the genome of human herpesvirus 6 encoding human cytomegalovirus immediate-early gene homologs and transactivating functions. J Virol. 1994 Feb;68(2):597–610. doi: 10.1128/jvi.68.2.597-610.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rawlinson W. D., Farrell H. E., Barrell B. G. Analysis of the complete DNA sequence of murine cytomegalovirus. J Virol. 1996 Dec;70(12):8833–8849. doi: 10.1128/jvi.70.12.8833-8849.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Stoddart C. A., Cardin R. D., Boname J. M., Manning W. C., Abenes G. B., Mocarski E. S. Peripheral blood mononuclear phagocytes mediate dissemination of murine cytomegalovirus. J Virol. 1994 Oct;68(10):6243–6253. doi: 10.1128/jvi.68.10.6243-6253.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Telford E. A., Watson M. S., Aird H. C., Perry J., Davison A. J. The DNA sequence of equine herpesvirus 2. J Mol Biol. 1995 Jun 9;249(3):520–528. doi: 10.1006/jmbi.1995.0314. [DOI] [PubMed] [Google Scholar]
  30. Thompson J. D., Higgins D. G., Gibson T. J. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994 Nov 11;22(22):4673–4680. doi: 10.1093/nar/22.22.4673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Thäle R., Lucin P., Schneider K., Eggers M., Koszinowski U. H. Identification and expression of a murine cytomegalovirus early gene coding for an Fc receptor. J Virol. 1994 Dec;68(12):7757–7765. doi: 10.1128/jvi.68.12.7757-7765.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Welch A. R., McGregor L. M., Gibson W. Cytomegalovirus homologs of cellular G protein-coupled receptor genes are transcribed. J Virol. 1991 Jul;65(7):3915–3918. doi: 10.1128/jvi.65.7.3915-3918.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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