Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1987 Feb;61(2):446–453. doi: 10.1128/jvi.61.2.446-453.1987

Primary structure and transcription of the genes coding for the two virion phosphoproteins pp65 and pp71 of human cytomegalovirus.

B Rüger, S Klages, B Walla, J Albrecht, B Fleckenstein, P Tomlinson, B Barrell
PMCID: PMC253968  PMID: 3027374

Abstract

Human cytomegalovirus contains a phosphorylated matrix protein of 65,000 apparent molecular weight (65K phosphoprotein; pp65) and a related phosphoprotein of 71,000 molecular weight (pp71). The 65K phosphoprotein is usually by far the most abundant structural component found in culture-grown purified virus particles. This study describes the precise mapping of the genes for both polypeptides, giving the entire nucleotide sequences and the exact positions of the respective transcripts. The 65K phosphoprotein is coded for by the 5'-terminal part of an abundant 4-kilobase (kb) mRNA. The 71K phosphoprotein corresponds to the single translational reading frame of a rare nonspliced 1.9-kb mRNA that is coterminal with the 4-kb transcript. The promoter for 4-kb mRNA appears to be unusual in structure; it does not contain a characteristic TATA sequence. The expression of antigenic epitopes from pp65 may allow improved serodiagnosis of human cytomegalovirus infections.

Full text

PDF
446

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bowden R. A., Sayers M., Flournoy N., Newton B., Banaji M., Thomas E. D., Meyers J. D. Cytomegalovirus immune globulin and seronegative blood products to prevent primary cytomegalovirus infection after marrow transplantation. N Engl J Med. 1986 Apr 17;314(16):1006–1010. doi: 10.1056/NEJM198604173141602. [DOI] [PubMed] [Google Scholar]
  2. Britt W. J., Auger D. Identification of a 65 000 dalton virion envelope protein of human cytomegalovirus. Virus Res. 1985 Dec;4(1):31–36. doi: 10.1016/0168-1702(85)90018-8. [DOI] [PubMed] [Google Scholar]
  3. Bzik D. J., Preston C. M. Analysis of DNA sequences which regulate the transcription of herpes simplex virus immediate early gene 3: DNA sequences required for enhancer-like activity and response to trans-activation by a virion polypeptide. Nucleic Acids Res. 1986 Jan 24;14(2):929–943. doi: 10.1093/nar/14.2.929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clark B. R., Zaia J. A., Balce-Directo L., Ting Y. P. Isolation and partial chemical characterization of a 64,000-dalton glycoprotein of human cytomegalovirus. J Virol. 1984 Jan;49(1):279–282. doi: 10.1128/jvi.49.1.279-282.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Davis M. G., Huang E. S. Nucleotide sequence of a human cytomegalovirus DNA fragment encoding a 67-kilodalton phosphorylated viral protein. J Virol. 1985 Oct;56(1):7–11. doi: 10.1128/jvi.56.1.7-11.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Davis M. G., Mar E. C., Wu Y. M., Huang E. S. Mapping and expression of a human cytomegalovirus major viral protein. J Virol. 1984 Oct;52(1):129–135. doi: 10.1128/jvi.52.1.129-135.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fleckenstein B., Müller I., Collins J. Cloning of the complete human cytomegalovirus genome in cosmids. Gene. 1982 Apr;18(1):39–46. doi: 10.1016/0378-1119(82)90054-3. [DOI] [PubMed] [Google Scholar]
  8. Geballe A. P., Leach F. S., Mocarski E. S. Regulation of cytomegalovirus late gene expression: gamma genes are controlled by posttranscriptional events. J Virol. 1986 Mar;57(3):864–874. doi: 10.1128/jvi.57.3.864-874.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gibson W. Protein counterparts of human and simian cytomegaloviruses. Virology. 1983 Jul 30;128(2):391–406. doi: 10.1016/0042-6822(83)90265-9. [DOI] [PubMed] [Google Scholar]
  10. Gibson W. Structural and nonstructural proteins of strain Colburn cytomegalovirus. Virology. 1981 Jun;111(2):516–537. doi: 10.1016/0042-6822(81)90354-8. [DOI] [PubMed] [Google Scholar]
  11. Irmiere A., Gibson W. Isolation and characterization of a noninfectious virion-like particle released from cells infected with human strains of cytomegalovirus. Virology. 1983 Oct 15;130(1):118–133. doi: 10.1016/0042-6822(83)90122-8. [DOI] [PubMed] [Google Scholar]
  12. Jahn G., Knust E., Schmolla H., Sarre T., Nelson J. A., McDougall J. K., Fleckenstein B. Predominant immediate-early transcripts of human cytomegalovirus AD 169. J Virol. 1984 Feb;49(2):363–370. doi: 10.1128/jvi.49.2.363-370.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Landini M. P., Re M. C., Mirolo G., Baldassarri B., La Placa M. Human immune response to cytomegalovirus structural polypeptides studied by immunoblotting. J Med Virol. 1985 Dec;17(4):303–311. doi: 10.1002/jmv.1890170403. [DOI] [PubMed] [Google Scholar]
  14. Mach M., Utz U., Fleckenstein B. Mapping of the major glycoprotein gene of human cytomegalovirus. J Gen Virol. 1986 Jul;67(Pt 7):1461–1467. doi: 10.1099/0022-1317-67-7-1461. [DOI] [PubMed] [Google Scholar]
  15. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  16. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  18. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  19. Nowak B., Gmeiner A., Sarnow P., Levine A. J., Fleckenstein B. Physical mapping of human cytomegalovirus genes: identification of DNA sequences coding for a virion phosphoprotein of 71 kDa and a viral 65-kDa polypeptide. Virology. 1984 Apr 15;134(1):91–102. doi: 10.1016/0042-6822(84)90275-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Pande H., Baak S. W., Riggs A. D., Clark B. R., Shively J. E., Zaia J. A. Cloning and physical mapping of a gene fragment coding for a 64-kilodalton major late antigen of human cytomegalovirus. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4965–4969. doi: 10.1073/pnas.81.15.4965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pellett P. E., McKnight J. L., Jenkins F. J., Roizman B. Nucleotide sequence and predicted amino acid sequence of a protein encoded in a small herpes simplex virus DNA fragment capable of trans-inducing alpha genes. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5870–5874. doi: 10.1073/pnas.82.17.5870. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rautmann G., Matthes H. W., Gait M. J., Breathnach R. Synthetic donor and acceptor splice sites function in an RNA polymerase B (II) transcription unit. EMBO J. 1984 Sep;3(9):2021–2028. doi: 10.1002/j.1460-2075.1984.tb02085.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  24. Roby C., Gibson W. Characterization of phosphoproteins and protein kinase activity of virions, noninfectious enveloped particles, and dense bodies of human cytomegalovirus. J Virol. 1986 Sep;59(3):714–727. doi: 10.1128/jvi.59.3.714-727.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  27. Staden R. A computer program to enter DNA gel reading data into a computer. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):499–503. doi: 10.1093/nar/12.1part2.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Staden R. Automation of the computer handling of gel reading data produced by the shotgun method of DNA sequencing. Nucleic Acids Res. 1982 Aug 11;10(15):4731–4751. doi: 10.1093/nar/10.15.4731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Talbot P., Almeida J. D. Human cytomegalovirus: purification of enveloped virions and dense bodies. J Gen Virol. 1977 Aug;36(2):345–349. doi: 10.1099/0022-1317-36-2-345. [DOI] [PubMed] [Google Scholar]
  30. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  31. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES