Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1991 Feb;65(2):649–656. doi: 10.1128/jvi.65.2.649-656.1991

The 68-kilodalton E1 protein of bovine papillomavirus is a DNA binding phosphoprotein which associates with the E2 transcriptional activator in vitro.

I L Blitz 1, L A Laimins 1
PMCID: PMC239803  PMID: 1846189

Abstract

The E1 open reading frame of bovine papillomavirus type 1 encodes factors necessary for extrachromosomal maintenance of the viral genome in transformed cells. To facilitate biochemical characterization of the gene products encoded by this open reading frame, we have expressed the full-length E1 protein in a baculovirus-insect cell system. This protein was found to be phosphorylated and localized to the nucleus of infected cells. The E1 protein alone has affinity for DNA but appears to lack specificity for viral sequences. In addition, we present evidence that the E1 protein interacts with the virally encoded transcriptional activator E2 in vitro. These results are consistent with a model in which the E1 protein, as part of a complex with E2, interacts with specific DNA sequences in the viral genome.

Full text

PDF
649

Images in this article

Selected References

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

  1. Androphy E. J., Lowy D. R., Schiller J. T. Bovine papillomavirus E2 trans-activating gene product binds to specific sites in papillomavirus DNA. Nature. 1987 Jan 1;325(6099):70–73. doi: 10.1038/325070a0. [DOI] [PubMed] [Google Scholar]
  2. Arthur A. K., Höss A., Fanning E. Expression of simian virus 40 T antigen in Escherichia coli: localization of T-antigen origin DNA-binding domain to within 129 amino acids. J Virol. 1988 Jun;62(6):1999–2006. doi: 10.1128/jvi.62.6.1999-2006.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berg L., Lusky M., Stenlund A., Botchan M. R. Repression of bovine papilloma virus replication is mediated by a virally encoded trans-acting factor. Cell. 1986 Aug 29;46(5):753–762. doi: 10.1016/0092-8674(86)90351-x. [DOI] [PubMed] [Google Scholar]
  4. Clertant P., Seif I. A common function for polyoma virus large-T and papillomavirus E1 proteins? Nature. 1984 Sep 20;311(5983):276–279. doi: 10.1038/311276a0. [DOI] [PubMed] [Google Scholar]
  5. Danos O., Giri I., Thierry F., Yaniv M. Papillomavirus genomes: sequences and consequences. J Invest Dermatol. 1984 Jul;83(1 Suppl):7s–11s. doi: 10.1111/1523-1747.ep12281115. [DOI] [PubMed] [Google Scholar]
  6. Dean F. B., Bullock P., Murakami Y., Wobbe C. R., Weissbach L., Hurwitz J. Simian virus 40 (SV40) DNA replication: SV40 large T antigen unwinds DNA containing the SV40 origin of replication. Proc Natl Acad Sci U S A. 1987 Jan;84(1):16–20. doi: 10.1073/pnas.84.1.16. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. DiMaio D. Nonsense mutation in open reading frame E2 of bovine papillomavirus DNA. J Virol. 1986 Feb;57(2):475–480. doi: 10.1128/jvi.57.2.475-480.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. DiMaio D., Settleman J. Bovine papillomavirus mutant temperature sensitive for transformation, replication and transactivation. EMBO J. 1988 Apr;7(4):1197–1204. doi: 10.1002/j.1460-2075.1988.tb02931.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Doorbar J., Campbell D., Grand R. J., Gallimore P. H. Identification of the human papilloma virus-1a E4 gene products. EMBO J. 1986 Feb;5(2):355–362. doi: 10.1002/j.1460-2075.1986.tb04219.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Groff D. E., Lancaster W. D. Genetic analysis of the 3' early region transformation and replication functions of bovine papillomavirus type 1. Virology. 1986 Apr 15;150(1):221–230. doi: 10.1016/0042-6822(86)90281-3. [DOI] [PubMed] [Google Scholar]
  12. Höss A., Moarefi I., Scheidtmann K. H., Cisek L. J., Corden J. L., Dornreiter I., Arthur A. K., Fanning E. Altered phosphorylation pattern of simian virus 40 T antigen expressed in insect cells by using a baculovirus vector. J Virol. 1990 Oct;64(10):4799–4807. doi: 10.1128/jvi.64.10.4799-4807.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kristie T. M., LeBowitz J. H., Sharp P. A. The octamer-binding proteins form multi-protein--DNA complexes with the HSV alpha TIF regulatory protein. EMBO J. 1989 Dec 20;8(13):4229–4238. doi: 10.1002/j.1460-2075.1989.tb08608.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kristie T. M., Roizman B. Host cell proteins bind to the cis-acting site required for virion-mediated induction of herpes simplex virus 1 alpha genes. Proc Natl Acad Sci U S A. 1987 Jan;84(1):71–75. doi: 10.1073/pnas.84.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lambert P. F., Howley P. M. Bovine papillomavirus type 1 E1 replication-defective mutants are altered in their transcriptional regulation. J Virol. 1988 Nov;62(11):4009–4015. doi: 10.1128/jvi.62.11.4009-4015.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Learned R. M., Learned T. K., Haltiner M. M., Tjian R. T. Human rRNA transcription is modulated by the coordinate binding of two factors to an upstream control element. Cell. 1986 Jun 20;45(6):847–857. doi: 10.1016/0092-8674(86)90559-3. [DOI] [PubMed] [Google Scholar]
  17. Luckow V. A., Summers M. D. High level expression of nonfused foreign genes with Autographa californica nuclear polyhedrosis virus expression vectors. Virology. 1989 May;170(1):31–39. doi: 10.1016/0042-6822(89)90348-6. [DOI] [PubMed] [Google Scholar]
  18. Lusky M., Botchan M. R. Genetic analysis of bovine papillomavirus type 1 trans-acting replication factors. J Virol. 1985 Mar;53(3):955–965. doi: 10.1128/jvi.53.3.955-965.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lusky M., Botchan M. R. Transient replication of bovine papilloma virus type 1 plasmids: cis and trans requirements. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3609–3613. doi: 10.1073/pnas.83.11.3609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. McBride A. A., Bolen J. B., Howley P. M. Phosphorylation sites of the E2 transcriptional regulatory proteins of bovine papillomavirus type 1. J Virol. 1989 Dec;63(12):5076–5085. doi: 10.1128/jvi.63.12.5076-5085.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. McKay R. D. Binding of a simian virus 40 T antigen-related protein to DNA. J Mol Biol. 1981 Jan 25;145(3):471–488. doi: 10.1016/0022-2836(81)90540-4. [DOI] [PubMed] [Google Scholar]
  22. McLean I. W., Nakane P. K. Periodate-lysine-paraformaldehyde fixative. A new fixation for immunoelectron microscopy. J Histochem Cytochem. 1974 Dec;22(12):1077–1083. doi: 10.1177/22.12.1077. [DOI] [PubMed] [Google Scholar]
  23. McVey D., Strauss M., Gluzman Y. Properties of the DNA-binding domain of the simian virus 40 large T antigen. Mol Cell Biol. 1989 Dec;9(12):5525–5536. doi: 10.1128/mcb.9.12.5525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Miller L. K. Baculoviruses as gene expression vectors. Annu Rev Microbiol. 1988;42:177–199. doi: 10.1146/annurev.mi.42.100188.001141. [DOI] [PubMed] [Google Scholar]
  25. Orth G., Jeanteur P., Croissant O. Evidence for and localization of vegetative viral DNA replication by autoradiographic detection of RNA-DNA hybrids in sections of tumors induced by Shope papilloma virus. Proc Natl Acad Sci U S A. 1971 Aug;68(8):1876–1880. doi: 10.1073/pnas.68.8.1876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Paucha E., Kalderon D., Harvey R. W., Smith A. E. Simian virus 40 origin DNA-binding domain on large T antigen. J Virol. 1986 Jan;57(1):50–64. doi: 10.1128/jvi.57.1.50-64.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Prives C. The replication functions of SV40 T antigen are regulated by phosphorylation. Cell. 1990 Jun 1;61(5):735–738. doi: 10.1016/0092-8674(90)90179-i. [DOI] [PubMed] [Google Scholar]
  28. Rabson M. S., Yee C., Yang Y. C., Howley P. M. Bovine papillomavirus type 1 3' early region transformation and plasmid maintenance functions. J Virol. 1986 Nov;60(2):626–634. doi: 10.1128/jvi.60.2.626-634.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rauscher F. J., 3rd, Voulalas P. J., Franza B. R., Jr, Curran T. Fos and Jun bind cooperatively to the AP-1 site: reconstitution in vitro. Genes Dev. 1988 Dec;2(12B):1687–1699. doi: 10.1101/gad.2.12b.1687. [DOI] [PubMed] [Google Scholar]
  30. Roberts J. M., Weintraub H. Negative control of DNA replication in composite SV40-bovine papilloma virus plasmids. Cell. 1986 Aug 29;46(5):741–752. doi: 10.1016/0092-8674(86)90350-8. [DOI] [PubMed] [Google Scholar]
  31. Santucci S., Androphy E. J., Bonne-Andréa C., Clertant P. Proteins encoded by the bovine papillomavirus E1 open reading frame: expression in heterologous systems and in virally transformed cells. J Virol. 1990 Dec;64(12):6027–6039. doi: 10.1128/jvi.64.12.6027-6039.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sarver N., Byrne J. C., Howley P. M. Transformation and replication in mouse cells of a bovine papillomavirus--pML2 plasmid vector that can be rescued in bacteria. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7147–7151. doi: 10.1073/pnas.79.23.7147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sarver N., Rabson M. S., Yang Y. C., Byrne J. C., Howley P. M. Localization and analysis of bovine papillomavirus type 1 transforming functions. J Virol. 1984 Nov;52(2):377–388. doi: 10.1128/jvi.52.2.377-388.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Schiller J. T., Kleiner E., Androphy E. J., Lowy D. R., Pfister H. Identification of bovine papillomavirus E1 mutants with increased transforming and transcriptional activity. J Virol. 1989 Apr;63(4):1775–1782. doi: 10.1128/jvi.63.4.1775-1782.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Seif I. Sequence homology between the large tumor antigen of polyoma viruses and the putative E1 protein of papilloma viruses. Virology. 1984 Oct 30;138(2):347–352. doi: 10.1016/0042-6822(84)90359-3. [DOI] [PubMed] [Google Scholar]
  36. Shapiro D. J., Sharp P. A., Wahli W. W., Keller M. J. A high-efficiency HeLa cell nuclear transcription extract. DNA. 1988 Jan-Feb;7(1):47–55. doi: 10.1089/dna.1988.7.47. [DOI] [PubMed] [Google Scholar]
  37. Simmons D. T. DNA-binding region of the simian virus 40 tumor antigen. J Virol. 1986 Mar;57(3):776–785. doi: 10.1128/jvi.57.3.776-785.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Simmons D. T. Geometry of the simian virus 40 large tumor antigen-DNA complex as probed by protease digestion. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2086–2090. doi: 10.1073/pnas.85.7.2086. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Spalholz B. A., Yang Y. C., Howley P. M. Transactivation of a bovine papilloma virus transcriptional regulatory element by the E2 gene product. Cell. 1985 Aug;42(1):183–191. doi: 10.1016/s0092-8674(85)80114-8. [DOI] [PubMed] [Google Scholar]
  40. Stahl H., Dröge P., Knippers R. DNA helicase activity of SV40 large tumor antigen. EMBO J. 1986 Aug;5(8):1939–1944. doi: 10.1002/j.1460-2075.1986.tb04447.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Stanley K. K., Luzio J. P. Construction of a new family of high efficiency bacterial expression vectors: identification of cDNA clones coding for human liver proteins. EMBO J. 1984 Jun;3(6):1429–1434. doi: 10.1002/j.1460-2075.1984.tb01988.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Strauss M., Argani P., Mohr I. J., Gluzman Y. Studies on the origin-specific DNA-binding domain of simian virus 40 large T antigen. J Virol. 1987 Oct;61(10):3326–3330. doi: 10.1128/jvi.61.10.3326-3330.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Sun S., Thorner L., Lentz M., MacPherson P., Botchan M. Identification of a 68-kilodalton nuclear ATP-binding phosphoprotein encoded by bovine papillomavirus type 1. J Virol. 1990 Oct;64(10):5093–5105. doi: 10.1128/jvi.64.10.5093-5105.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Thorner L., Bucay N., Choe J., Botchan M. The product of the bovine papillomavirus type 1 modulator gene (M) is a phosphoprotein. J Virol. 1988 Jul;62(7):2474–2482. doi: 10.1128/jvi.62.7.2474-2482.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tjian R., Robbins A. Enzymatic activities associated with a purified simian virus 40 T antigen-related protein. Proc Natl Acad Sci U S A. 1979 Feb;76(2):610–614. doi: 10.1073/pnas.76.2.610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Tjian R. The binding site on SV40 DNA for a T antigen-related protein. Cell. 1978 Jan;13(1):165–179. doi: 10.1016/0092-8674(78)90147-2. [DOI] [PubMed] [Google Scholar]
  47. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Tratner I., De Togni P., Sassone-Corsi P., Verma I. M. Characterization and purification of human fos protein generated in insect cells with a baculoviral expression vector. J Virol. 1990 Feb;64(2):499–508. doi: 10.1128/jvi.64.2.499-508.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wun-Kim K., Simmons D. T. Mapping of helicase and helicase substrate-binding domains on simian virus 40 large T antigen. J Virol. 1990 May;64(5):2014–2020. doi: 10.1128/jvi.64.5.2014-2020.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES