Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1997 May;71(5):3853–3863. doi: 10.1128/jvi.71.5.3853-3863.1997

Functional interactions between papillomavirus E1 and E2 proteins.

M Berg 1, A Stenlund 1
PMCID: PMC191536  PMID: 9094661

Abstract

DNA replication of papillomaviruses requires the viral E1 and E2 proteins. These proteins bind cooperatively to the viral origin of replication (ori), which contains binding sites for both proteins, forming an E1-E2-ori complex which is essential for initiation of DNA replication. To map the domains in E2 that are involved in the interaction with E1, we have used chimeric bovine papillomavirus (BPV)/human papillomavirus type 11 (HPV-11) E2 proteins. The results from this study show that both the DNA binding domain and the transactivation domain from BPV E2 independently can interact with BPV E1. However, the roles of these two interactions are different: the interaction between E1 and the activation domain of E2 is necessary and sufficient for cooperativity in binding and for DNA replication; the interaction between E1 and the DNA binding domain of E2 is required only when the binding sites for E1 and E2 are adjacent to each other, and the function of this interaction appears to be to facilitate the interaction between E1 and the transactivation domain of E2. These results indicate that the cooperative binding of E1 and E2 to the BPV ori takes place via a novel two-stage mechanism where one interaction serves as a trigger for the formation of the second, productive, interaction between the two proteins.

Full Text

The Full Text of this article is available as a PDF (778.5 KB).

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. Arndt K., Fink G. R. GCN4 protein, a positive transcription factor in yeast, binds general control promoters at all 5' TGACTC 3' sequences. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8516–8520. doi: 10.1073/pnas.83.22.8516. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bennett-Cook E. R., Hassell J. A. Activation of polyomavirus DNA replication by yeast GAL4 is dependent on its transcriptional activation domains. EMBO J. 1991 Apr;10(4):959–969. doi: 10.1002/j.1460-2075.1991.tb08030.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Benson J. D., Howley P. M. Amino-terminal domains of the bovine papillomavirus type 1 E1 and E2 proteins participate in complex formation. J Virol. 1995 Jul;69(7):4364–4372. doi: 10.1128/jvi.69.7.4364-4372.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blitz I. L., Laimins L. A. The 68-kilodalton E1 protein of bovine papillomavirus is a DNA binding phosphoprotein which associates with the E2 transcriptional activator in vitro. J Virol. 1991 Feb;65(2):649–656. doi: 10.1128/jvi.65.2.649-656.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bosher J., Robinson E. C., Hay R. T. Interactions between the adenovirus type 2 DNA polymerase and the DNA binding domain of nuclear factor I. New Biol. 1990 Dec;2(12):1083–1090. [PubMed] [Google Scholar]
  7. Brokaw J. L., Blanco M., McBride A. A. Amino acids critical for the functions of the bovine papillomavirus type 1 E2 transactivator. J Virol. 1996 Jan;70(1):23–29. doi: 10.1128/jvi.70.1.23-29.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Challberg M. D., Kelly T. J. Animal virus DNA replication. Annu Rev Biochem. 1989;58:671–717. doi: 10.1146/annurev.bi.58.070189.003323. [DOI] [PubMed] [Google Scholar]
  9. Cheng L. Z., Workman J. L., Kingston R. E., Kelly T. J. Regulation of DNA replication in vitro by the transcriptional activation domain of GAL4-VP16. Proc Natl Acad Sci U S A. 1992 Jan 15;89(2):589–593. doi: 10.1073/pnas.89.2.589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cheng L., Kelly T. J. Transcriptional activator nuclear factor I stimulates the replication of SV40 minichromosomes in vivo and in vitro. Cell. 1989 Nov 3;59(3):541–551. doi: 10.1016/0092-8674(89)90037-8. [DOI] [PubMed] [Google Scholar]
  11. Chiang C. M., Ustav M., Stenlund A., Ho T. F., Broker T. R., Chow L. T. Viral E1 and E2 proteins support replication of homologous and heterologous papillomaviral origins. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):5799–5803. doi: 10.1073/pnas.89.13.5799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. DePamphilis M. L. Eukaryotic DNA replication: anatomy of an origin. Annu Rev Biochem. 1993;62:29–63. doi: 10.1146/annurev.bi.62.070193.000333. [DOI] [PubMed] [Google Scholar]
  13. Del Vecchio A. M., Romanczuk H., Howley P. M., Baker C. C. Transient replication of human papillomavirus DNAs. J Virol. 1992 Oct;66(10):5949–5958. doi: 10.1128/jvi.66.10.5949-5958.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ferguson M. K., Botchan M. R. Genetic analysis of the activation domain of bovine papillomavirus protein E2: its role in transcription and replication. J Virol. 1996 Jul;70(7):4193–4199. doi: 10.1128/jvi.70.7.4193-4199.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gillette T. G., Lusky M., Borowiec J. A. Induction of structural changes in the bovine papillomavirus type 1 origin of replication by the viral E1 and E2 proteins. Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):8846–8850. doi: 10.1073/pnas.91.19.8846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Giri I., Yaniv M. Structural and mutational analysis of E2 trans-activating proteins of papillomaviruses reveals three distinct functional domains. EMBO J. 1988 Sep;7(9):2823–2829. doi: 10.1002/j.1460-2075.1988.tb03138.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Grossel M. J., Sverdrup F., Breiding D. E., Androphy E. J. Transcriptional activation function is not required for stimulation of DNA replication by bovine papillomavirus type 1 E2. J Virol. 1996 Oct;70(10):7264–7269. doi: 10.1128/jvi.70.10.7264-7269.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Guo Z. S., DePamphilis M. L. Specific transcription factors stimulate simian virus 40 and polyomavirus origins of DNA replication. Mol Cell Biol. 1992 Jun;12(6):2514–2524. doi: 10.1128/mcb.12.6.2514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hawley-Nelson P., Androphy E. J., Lowy D. R., Schiller J. T. The specific DNA recognition sequence of the bovine papillomavirus E2 protein is an E2-dependent enhancer. EMBO J. 1988 Feb;7(2):525–531. doi: 10.1002/j.1460-2075.1988.tb02841.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. He Z., Brinton B. T., Greenblatt J., Hassell J. A., Ingles C. J. The transactivator proteins VP16 and GAL4 bind replication factor A. Cell. 1993 Jun 18;73(6):1223–1232. doi: 10.1016/0092-8674(93)90650-f. [DOI] [PubMed] [Google Scholar]
  21. Heintz N. H. Transcription factors and the control of DNA replication. Curr Opin Cell Biol. 1992 Jun;4(3):459–467. doi: 10.1016/0955-0674(92)90012-2. [DOI] [PubMed] [Google Scholar]
  22. Hibma M. H., Raj K., Ely S. J., Stanley M., Crawford L. The interaction between human papillomavirus type 16 E1 and E2 proteins is blocked by an antibody to the N-terminal region of E2. Eur J Biochem. 1995 Apr 15;229(2):517–525. doi: 10.1111/j.1432-1033.1995.0517k.x. [DOI] [PubMed] [Google Scholar]
  23. Holt S. E., Schuller G., Wilson V. G. DNA binding specificity of the bovine papillomavirus E1 protein is determined by sequences contained within an 18-base-pair inverted repeat element at the origin of replication. J Virol. 1994 Feb;68(2):1094–1102. doi: 10.1128/jvi.68.2.1094-1102.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Holt S. E., Wilson V. G. Mutational analysis of the 18-base-pair inverted repeat element at the bovine papillomavirus origin of replication: identification of critical sequences for E1 binding and in vivo replication. J Virol. 1995 Oct;69(10):6525–6532. doi: 10.1128/jvi.69.10.6525-6532.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Hope I. A., Struhl K. Functional dissection of a eukaryotic transcriptional activator protein, GCN4 of yeast. Cell. 1986 Sep 12;46(6):885–894. doi: 10.1016/0092-8674(86)90070-x. [DOI] [PubMed] [Google Scholar]
  26. Hubbert N. L., Schiller J. T., Lowy D. R., Androphy E. J. Bovine papilloma virus-transformed cells contain multiple E2 proteins. Proc Natl Acad Sci U S A. 1988 Aug;85(16):5864–5868. doi: 10.1073/pnas.85.16.5864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kadonaga J. T. Purification of sequence-specific binding proteins by DNA affinity chromatography. Methods Enzymol. 1991;208:10–23. doi: 10.1016/0076-6879(91)08004-2. [DOI] [PubMed] [Google Scholar]
  28. Lambert P. F., Hubbert N. L., Howley P. M., Schiller J. T. Genetic assignment of multiple E2 gene products in bovine papillomavirus-transformed cells. J Virol. 1989 Jul;63(7):3151–3154. doi: 10.1128/jvi.63.7.3151-3154.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Li R., Botchan M. R. Acidic transcription factors alleviate nucleosome-mediated repression of DNA replication of bovine papillomavirus type 1. Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7051–7055. doi: 10.1073/pnas.91.15.7051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Li R., Botchan M. R. The acidic transcriptional activation domains of VP16 and p53 bind the cellular replication protein A and stimulate in vitro BPV-1 DNA replication. Cell. 1993 Jun 18;73(6):1207–1221. doi: 10.1016/0092-8674(93)90649-b. [DOI] [PubMed] [Google Scholar]
  31. Lusky M., Fontane E. Formation of the complex of bovine papillomavirus E1 and E2 proteins is modulated by E2 phosphorylation and depends upon sequences within the carboxyl terminus of E1. Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6363–6367. doi: 10.1073/pnas.88.14.6363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Lusky M., Hurwitz J., Seo Y. S. The bovine papillomavirus E2 protein modulates the assembly of but is not stably maintained in a replication-competent multimeric E1-replication origin complex. Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):8895–8899. doi: 10.1073/pnas.91.19.8895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. McBride A. A., Romanczuk H., Howley P. M. The papillomavirus E2 regulatory proteins. J Biol Chem. 1991 Oct 5;266(28):18411–18414. [PubMed] [Google Scholar]
  34. 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]
  35. Mohr I. J., Clark R., Sun S., Androphy E. J., MacPherson P., Botchan M. R. Targeting the E1 replication protein to the papillomavirus origin of replication by complex formation with the E2 transactivator. Science. 1990 Dec 21;250(4988):1694–1699. doi: 10.1126/science.2176744. [DOI] [PubMed] [Google Scholar]
  36. Mul Y. M., Van der Vliet P. C. Nuclear factor I enhances adenovirus DNA replication by increasing the stability of a preinitiation complex. EMBO J. 1992 Feb;11(2):751–760. doi: 10.1002/j.1460-2075.1992.tb05108.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Mul Y. M., Verrijzer C. P., van der Vliet P. C. Transcription factors NFI and NFIII/oct-1 function independently, employing different mechanisms to enhance adenovirus DNA replication. J Virol. 1990 Nov;64(11):5510–5518. doi: 10.1128/jvi.64.11.5510-5518.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Piccini A., Storey A., Massimi P., Banks L. Mutations in the human papillomavirus type 16 E2 protein identify multiple regions of the protein involved in binding to E1. J Gen Virol. 1995 Nov;76(Pt 11):2909–2913. doi: 10.1099/0022-1317-76-11-2909. [DOI] [PubMed] [Google Scholar]
  39. Sakai H., Yasugi T., Benson J. D., Dowhanick J. J., Howley P. M. Targeted mutagenesis of the human papillomavirus type 16 E2 transactivation domain reveals separable transcriptional activation and DNA replication functions. J Virol. 1996 Mar;70(3):1602–1611. doi: 10.1128/jvi.70.3.1602-1611.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Schägger H., von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987 Nov 1;166(2):368–379. doi: 10.1016/0003-2697(87)90587-2. [DOI] [PubMed] [Google Scholar]
  41. Sedman J., Stenlund A. Co-operative interaction between the initiator E1 and the transcriptional activator E2 is required for replicator specific DNA replication of bovine papillomavirus in vivo and in vitro. EMBO J. 1995 Dec 15;14(24):6218–6228. doi: 10.1002/j.1460-2075.1995.tb00312.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sedman J., Stenlund A. The initiator protein E1 binds to the bovine papillomavirus origin of replication as a trimeric ring-like structure. EMBO J. 1996 Sep 16;15(18):5085–5092. [PMC free article] [PubMed] [Google Scholar]
  43. Sedman T., Sedman J., Stenlund A. Binding of the E1 and E2 proteins to the origin of replication of bovine papillomavirus. J Virol. 1997 Apr;71(4):2887–2896. doi: 10.1128/jvi.71.4.2887-2896.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Seo Y. S., Müller F., Lusky M., Gibbs E., Kim H. Y., Phillips B., Hurwitz J. Bovine papilloma virus (BPV)-encoded E2 protein enhances binding of E1 protein to the BPV replication origin. Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2865–2869. doi: 10.1073/pnas.90.7.2865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Seo Y. S., Müller F., Lusky M., Hurwitz J. Bovine papilloma virus (BPV)-encoded E1 protein contains multiple activities required for BPV DNA replication. Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):702–706. doi: 10.1073/pnas.90.2.702. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Spalholz B. A., McBride A. A., Sarafi T., Quintero J. Binding of bovine papillomavirus E1 to the origin is not sufficient for DNA replication. Virology. 1993 Mar;193(1):201–212. doi: 10.1006/viro.1993.1116. [DOI] [PubMed] [Google Scholar]
  47. 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]
  48. Stillman B. Initiation of eukaryotic DNA replication in vitro. Annu Rev Cell Biol. 1989;5:197–245. doi: 10.1146/annurev.cb.05.110189.001213. [DOI] [PubMed] [Google Scholar]
  49. Storey A., Piccini A., Massimi P., Bouvard V., Banks L. Mutations in the human papillomavirus type 16 E2 protein identify a region of the protein involved in binding to E1 protein. J Gen Virol. 1995 Apr;76(Pt 4):819–826. doi: 10.1099/0022-1317-76-4-819. [DOI] [PubMed] [Google Scholar]
  50. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  51. Tanaka M., Herr W. Differential transcriptional activation by Oct-1 and Oct-2: interdependent activation domains induce Oct-2 phosphorylation. Cell. 1990 Feb 9;60(3):375–386. doi: 10.1016/0092-8674(90)90589-7. [DOI] [PubMed] [Google Scholar]
  52. Thorner L. K., Lim D. A., Botchan M. R. DNA-binding domain of bovine papillomavirus type 1 E1 helicase: structural and functional aspects. J Virol. 1993 Oct;67(10):6000–6014. doi: 10.1128/jvi.67.10.6000-6014.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Ustav E., Ustav M., Szymanski P., Stenlund A. The bovine papillomavirus origin of replication requires a binding site for the E2 transcriptional activator. Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):898–902. doi: 10.1073/pnas.90.3.898. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Ustav M., Stenlund A. Transient replication of BPV-1 requires two viral polypeptides encoded by the E1 and E2 open reading frames. EMBO J. 1991 Feb;10(2):449–457. doi: 10.1002/j.1460-2075.1991.tb07967.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Ustav M., Ustav E., Szymanski P., Stenlund A. Identification of the origin of replication of bovine papillomavirus and characterization of the viral origin recognition factor E1. EMBO J. 1991 Dec;10(13):4321–4329. doi: 10.1002/j.1460-2075.1991.tb05010.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Wilson V. G., Ludes-Meyers J. A bovine papillomavirus E1-related protein binds specifically to bovine papillomavirus DNA. J Virol. 1991 Oct;65(10):5314–5322. doi: 10.1128/jvi.65.10.5314-5322.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Winokur P. L., McBride A. A. Separation of the transcriptional activation and replication functions of the bovine papillomavirus-1 E2 protein. EMBO J. 1992 Nov;11(11):4111–4118. doi: 10.1002/j.1460-2075.1992.tb05504.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Winokur P. L., McBride A. A. The transactivation and DNA binding domains of the BPV-1 E2 protein have different roles in cooperative origin binding with the E1 protein. Virology. 1996 Jul 1;221(1):44–53. doi: 10.1006/viro.1996.0351. [DOI] [PubMed] [Google Scholar]
  59. Yang L., Li R., Mohr I. J., Clark R., Botchan M. R. Activation of BPV-1 replication in vitro by the transcription factor E2. Nature. 1991 Oct 17;353(6345):628–632. doi: 10.1038/353628a0. [DOI] [PubMed] [Google Scholar]
  60. Yang L., Mohr I., Fouts E., Lim D. A., Nohaile M., Botchan M. The E1 protein of bovine papilloma virus 1 is an ATP-dependent DNA helicase. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5086–5090. doi: 10.1073/pnas.90.11.5086. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Yang L., Mohr I., Li R., Nottoli T., Sun S., Botchan M. Transcription factor E2 regulates BPV-1 DNA replication in vitro by direct protein-protein interaction. Cold Spring Harb Symp Quant Biol. 1991;56:335–346. doi: 10.1101/sqb.1991.056.01.040. [DOI] [PubMed] [Google Scholar]

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

RESOURCES