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. 1994 Sep;68(9):6097–6102. doi: 10.1128/jvi.68.9.6097-6102.1994

Papilloma formation by cottontail rabbit papillomavirus requires E1 and E2 regulatory genes in addition to E6 and E7 transforming genes.

X Wu 1, W Xiao 1, J L Brandsma 1
PMCID: PMC237021  PMID: 8057490

Abstract

The present study used the cottontail rabbit papillomavirus DNA-rabbit system to evaluate whether the regulatory genes E1 and E2 and the transforming gene E6 are required for papilloma formation. Frameshift mutations were generated in the individual genes in the context of a full-length cottontail rabbit papillomavirus genome, and the mutant DNAs were intradermally inoculated into domestic rabbits. None of the mutants induced papillomas. Marker rescue experiments confirmed that the defects were due to mutations that we deliberately introduced. Marker rescue also confirmed our previous report that the upstream region of E7 around position 9 was critical for papilloma induction. These results demonstrate that the E1 and E2 regulatory genes as well as the E6 and E7 transforming genes are each required for papilloma formation. Each gene may provide molecular targets for therapeutic intervention.

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Selected References

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  1. Arbeit J. M., Münger K., Howley P. M., Hanahan D. Neuroepithelial carcinomas in mice transgenic with human papillomavirus type 16 E6/E7 ORFs. Am J Pathol. 1993 Apr;142(4):1187–1197. [PMC free article] [PubMed] [Google Scholar]
  2. Barbosa M. S., Wettstein F. O. Transcription of the cottontail rabbit papillomavirus early region and identification of two E6 polypeptides in COS-7 cells. J Virol. 1987 Sep;61(9):2938–2942. doi: 10.1128/jvi.61.9.2938-2942.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brandsma J. L., Xiao W. Infectious virus replication in papillomas induced by molecularly cloned cottontail rabbit papillomavirus DNA. J Virol. 1993 Jan;67(1):567–571. doi: 10.1128/jvi.67.1.567-571.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brandsma J. L., Yang Z. H., Barthold S. W., Johnson E. A. Use of a rapid, efficient inoculation method to induce papillomas by cottontail rabbit papillomavirus DNA shows that the E7 gene is required. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4816–4820. doi: 10.1073/pnas.88.11.4816. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brandsma J. L., Yang Z. H., DiMaio D., Barthold S. W., Johnson E., Xiao W. The putative E5 open reading frame of cottontail rabbit papillomavirus is dispensable for papilloma formation in domestic rabbits. J Virol. 1992 Oct;66(10):6204–6207. doi: 10.1128/jvi.66.10.6204-6207.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chiang C. M., Broker T. R., Chow L. T. Properties of bovine papillomavirus E1 mutants. Virology. 1992 Dec;191(2):964–967. doi: 10.1016/0042-6822(92)90273-r. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Crook T., Morgenstern J. P., Crawford L., Banks L. Continued expression of HPV-16 E7 protein is required for maintenance of the transformed phenotype of cells co-transformed by HPV-16 plus EJ-ras. EMBO J. 1989 Feb;8(2):513–519. doi: 10.1002/j.1460-2075.1989.tb03405.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Defeo-Jones D., Vuocolo G. A., Haskell K. M., Hanobik M. G., Kiefer D. M., McAvoy E. M., Ivey-Hoyle M., Brandsma J. L., Oliff A., Jones R. E. Papillomavirus E7 protein binding to the retinoblastoma protein is not required for viral induction of warts. J Virol. 1993 Feb;67(2):716–725. doi: 10.1128/jvi.67.2.716-725.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. DiMaio D. Transforming activity of bovine and human papillomaviruses in cultured cells. Adv Cancer Res. 1991;56:133–159. doi: 10.1016/s0065-230x(08)60480-7. [DOI] [PubMed] [Google Scholar]
  12. Dong G., Broker T. R., Chow L. T. Human papillomavirus type 11 E2 proteins repress the homologous E6 promoter by interfering with the binding of host transcription factors to adjacent elements. J Virol. 1994 Feb;68(2):1115–1127. doi: 10.1128/jvi.68.2.1115-1127.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dyson N., Howley P. M., Münger K., Harlow E. The human papilloma virus-16 E7 oncoprotein is able to bind to the retinoblastoma gene product. Science. 1989 Feb 17;243(4893):934–937. doi: 10.1126/science.2537532. [DOI] [PubMed] [Google Scholar]
  14. Friedman J. M., Fialkow P. J. Viral "tumorigenesis" in man: cell markers in condylomata acuminata. Int J Cancer. 1976 Jan 15;17(1):57–61. doi: 10.1002/ijc.2910170109. [DOI] [PubMed] [Google Scholar]
  15. Georges E., Pehau-Arnaudet G., Orth G. Molecular and biological characterization of cottontail rabbit papillomavirus variant DNA sequences integrated in the VX7 carcinoma. Virology. 1992 Feb;186(2):750–759. doi: 10.1016/0042-6822(92)90042-n. [DOI] [PubMed] [Google Scholar]
  16. Giri I., Danos O., Yaniv M. Genomic structure of the cottontail rabbit (Shope) papillomavirus. Proc Natl Acad Sci U S A. 1985 Mar;82(6):1580–1584. doi: 10.1073/pnas.82.6.1580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Giri I., Yaniv M. Study of the E2 gene product of the cottontail rabbit papillomavirus reveals a common mechanism of transactivation among papillomaviruses. J Virol. 1988 May;62(5):1573–1581. doi: 10.1128/jvi.62.5.1573-1581.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Griep A. E., Herber R., Jeon S., Lohse J. K., Dubielzig R. R., Lambert P. F. Tumorigenicity by human papillomavirus type 16 E6 and E7 in transgenic mice correlates with alterations in epithelial cell growth and differentiation. J Virol. 1993 Mar;67(3):1373–1384. doi: 10.1128/jvi.67.3.1373-1384.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ham J., Dostatni N., Gauthier J. M., Yaniv M. The papillomavirus E2 protein: a factor with many talents. Trends Biochem Sci. 1991 Nov;16(11):440–444. doi: 10.1016/0968-0004(91)90172-r. [DOI] [PubMed] [Google Scholar]
  21. Haskell K. M., Vuocolo G. A., Defeo-Jones D., Jones R. E., Ivey-Hoyle M. Comparison of the binding of the human papillomavirus type 16 and cottontail rabbit papillomavirus E7 proteins to the retinoblastoma gene product. J Gen Virol. 1993 Jan;74(Pt 1):115–119. doi: 10.1099/0022-1317-74-1-115. [DOI] [PubMed] [Google Scholar]
  22. Hawley-Nelson P., Vousden K. H., Hubbert N. L., Lowy D. R., Schiller J. T. HPV16 E6 and E7 proteins cooperate to immortalize human foreskin keratinocytes. EMBO J. 1989 Dec 1;8(12):3905–3910. doi: 10.1002/j.1460-2075.1989.tb08570.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hudson J. B., Bedell M. A., McCance D. J., Laiminis L. A. Immortalization and altered differentiation of human keratinocytes in vitro by the E6 and E7 open reading frames of human papillomavirus type 18. J Virol. 1990 Feb;64(2):519–526. doi: 10.1128/jvi.64.2.519-526.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kondoh G., Murata Y., Aozasa K., Yutsudo M., Hakura A. Very high incidence of germ cell tumorigenesis (seminomagenesis) in human papillomavirus type 16 transgenic mice. J Virol. 1991 Jun;65(6):3335–3339. doi: 10.1128/jvi.65.6.3335-3339.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kreider J. W., Bartlett G. L. The Shope papilloma-carcinoma complex of rabbits: a model system of neoplastic progression and spontaneous regression. Adv Cancer Res. 1981;35:81–110. doi: 10.1016/s0065-230x(08)60909-4. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Lambert P. F., Pan H., Pitot H. C., Liem A., Jackson M., Griep A. E. Epidermal cancer associated with expression of human papillomavirus type 16 E6 and E7 oncogenes in the skin of transgenic mice. Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5583–5587. doi: 10.1073/pnas.90.12.5583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lambert P. F. Papillomavirus DNA replication. J Virol. 1991 Jul;65(7):3417–3420. doi: 10.1128/jvi.65.7.3417-3420.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Meyers C., Harry J., Lin Y. L., Wettstein F. O. Identification of three transforming proteins encoded by cottontail rabbit papillomavirus. J Virol. 1992 Mar;66(3):1655–1664. doi: 10.1128/jvi.66.3.1655-1664.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Meyers C., Wettstein F. O. The late region differentially regulates the in vitro transformation by cottontail rabbit papillomavirus DNA in different cell types. Virology. 1991 Apr;181(2):637–646. doi: 10.1016/0042-6822(91)90897-k. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Mounts P., Shah K. V. Respiratory papillomatosis: etiological relation to genital tract papillomaviruses. Prog Med Virol. 1984;29:90–114. [PubMed] [Google Scholar]
  34. Münger K., Phelps W. C., Bubb V., Howley P. M., Schlegel R. The E6 and E7 genes of the human papillomavirus type 16 together are necessary and sufficient for transformation of primary human keratinocytes. J Virol. 1989 Oct;63(10):4417–4421. doi: 10.1128/jvi.63.10.4417-4421.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Nasseri M., Meyers C., Wettstein F. O. Genetic analysis of CRPV pathogenesis: the L1 open reading frame is dispensable for cellular transformation but is required for papilloma formation. Virology. 1989 May;170(1):321–325. doi: 10.1016/0042-6822(89)90388-7. [DOI] [PubMed] [Google Scholar]
  36. Nishikawa T., Yamashita T., Yamada T., Kobayashi H., Ohkawara A., Fujinaga K. Tumorigenic transformation of primary rat embryonal fibroblasts by human papillomavirus type 8 E7 gene in collaboration with the activated H-ras gene. Jpn J Cancer Res. 1991 Dec;82(12):1340–1343. doi: 10.1111/j.1349-7006.1991.tb01802.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Peng X., Olson R. O., Christian C. B., Lang C. M., Kreider J. W. Papillomas and carcinomas in transgenic rabbits carrying EJ-ras DNA and cottontail rabbit papillomavirus DNA. J Virol. 1993 Mar;67(3):1698–1701. doi: 10.1128/jvi.67.3.1698-1701.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Phelps W. C., Yee C. L., Münger K., Howley P. M. The human papillomavirus type 16 E7 gene encodes transactivation and transformation functions similar to those of adenovirus E1A. Cell. 1988 May 20;53(4):539–547. doi: 10.1016/0092-8674(88)90570-3. [DOI] [PubMed] [Google Scholar]
  39. Romanczuk H., Howley P. M. Disruption of either the E1 or the E2 regulatory gene of human papillomavirus type 16 increases viral immortalization capacity. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):3159–3163. doi: 10.1073/pnas.89.7.3159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sang B. C., Barbosa M. S. Increased E6/E7 transcription in HPV 18-immortalized human keratinocytes results from inactivation of E2 and additional cellular events. Virology. 1992 Aug;189(2):448–455. doi: 10.1016/0042-6822(92)90568-a. [DOI] [PubMed] [Google Scholar]
  41. Scheffner M., Werness B. A., Huibregtse J. M., Levine A. J., Howley P. M. The E6 oncoprotein encoded by human papillomavirus types 16 and 18 promotes the degradation of p53. Cell. 1990 Dec 21;63(6):1129–1136. doi: 10.1016/0092-8674(90)90409-8. [DOI] [PubMed] [Google Scholar]
  42. 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]
  43. Sedman S. A., Barbosa M. S., Vass W. C., Hubbert N. L., Haas J. A., Lowy D. R., Schiller J. T. The full-length E6 protein of human papillomavirus type 16 has transforming and trans-activating activities and cooperates with E7 to immortalize keratinocytes in culture. J Virol. 1991 Sep;65(9):4860–4866. doi: 10.1128/jvi.65.9.4860-4866.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. 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]
  45. Stoler M. H., Whitbeck A., Wolinsky S. M., Broker T. R., Chow L. T., Howett M. K., Kreider J. W. Infectious cycle of human papillomavirus type 11 in human foreskin xenografts in nude mice. J Virol. 1990 Jul;64(7):3310–3318. doi: 10.1128/jvi.64.7.3310-3318.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Stoler M. H., Wolinsky S. M., Whitbeck A., Broker T. R., Chow L. T. Differentiation-linked human papillomavirus types 6 and 11 transcription in genital condylomata revealed by in situ hybridization with message-specific RNA probes. Virology. 1989 Sep;172(1):331–340. doi: 10.1016/0042-6822(89)90135-9. [DOI] [PubMed] [Google Scholar]
  47. Szymanski P., Stenlund A. Regulation of early gene expression from the bovine papillomavirus genome in transiently transfected C127 cells. J Virol. 1991 Nov;65(11):5710–5720. doi: 10.1128/jvi.65.11.5710-5720.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Takami Y., Sasagawa T., Sudiro T. M., Yutsudo M., Hakura A. Determination of the functional difference between human papillomavirus type 6 and 16 E7 proteins by their 30 N-terminal amino acid residues. Virology. 1992 Feb;186(2):489–495. doi: 10.1016/0042-6822(92)90014-g. [DOI] [PubMed] [Google Scholar]
  49. Thierry F., Howley P. M. Functional analysis of E2-mediated repression of the HPV18 P105 promoter. New Biol. 1991 Jan;3(1):90–100. [PubMed] [Google Scholar]
  50. 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]
  51. 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]
  52. Watanabe S., Sato H., Komiyama N., Kanda T., Yoshiike K. The E7 functions of human papillomaviruses in rat 3Y1 cells. Virology. 1992 Mar;187(1):107–114. doi: 10.1016/0042-6822(92)90299-5. [DOI] [PubMed] [Google Scholar]
  53. Werness B. A., Levine A. J., Howley P. M. Association of human papillomavirus types 16 and 18 E6 proteins with p53. Science. 1990 Apr 6;248(4951):76–79. doi: 10.1126/science.2157286. [DOI] [PubMed] [Google Scholar]
  54. Wu E. W., Clemens K. E., Heck D. V., Münger K. The human papillomavirus E7 oncoprotein and the cellular transcription factor E2F bind to separate sites on the retinoblastoma tumor suppressor protein. J Virol. 1993 Apr;67(4):2402–2407. doi: 10.1128/jvi.67.4.2402-2407.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. 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]
  56. 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]

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