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. 1992 Jun;66(6):3941–3945. doi: 10.1128/jvi.66.6.3941-3945.1992

Mechanism of action of the papillomavirus E2 repressor: repression in the absence of DNA binding.

J Barsoum 1, S S Prakash 1, P Han 1, E J Androphy 1
PMCID: PMC241186  PMID: 1316493

Abstract

Repression of papillomavirus E2-dependent gene expression was studied by using transient transfections into mouse embryo fibroblast cells. Cotransfection of a gene corresponding to the naturally occurring repressor E2-TR along with the full-length E2 gene resulted in up to 98% repression of E2-dependent reporter gene expression. A series of E2 DNA-binding domain mutants were transferred into the E2-TR form and characterized for their ability to repress E2-dependent transactivation. All mutants which were defective for DNA binding but were dimerization competent repressed E2 transactivation as well or nearly as well as the wild-type repressor. E2 mutants which lacked dimerization activity repressed transactivation poorly or not at all. These results indicate that the E2 repressor can inhibit transcription, in the absence of DNA binding, by forming heterodimers with full-length E2.

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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. Benezra R., Davis R. L., Lockshon D., Turner D. L., Weintraub H. The protein Id: a negative regulator of helix-loop-helix DNA binding proteins. Cell. 1990 Apr 6;61(1):49–59. doi: 10.1016/0092-8674(90)90214-y. [DOI] [PubMed] [Google Scholar]
  3. Cate R. L., Mattaliano R. J., Hession C., Tizard R., Farber N. M., Cheung A., Ninfa E. G., Frey A. Z., Gash D. J., Chow E. P. Isolation of the bovine and human genes for Müllerian inhibiting substance and expression of the human gene in animal cells. Cell. 1986 Jun 6;45(5):685–698. doi: 10.1016/0092-8674(86)90783-x. [DOI] [PubMed] [Google Scholar]
  4. Choe J., Vaillancourt P., Stenlund A., Botchan M. Bovine papillomavirus type 1 encodes two forms of a transcriptional repressor: structural and functional analysis of new viral cDNAs. J Virol. 1989 Apr;63(4):1743–1755. doi: 10.1128/jvi.63.4.1743-1755.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chu G., Hayakawa H., Berg P. Electroporation for the efficient transfection of mammalian cells with DNA. Nucleic Acids Res. 1987 Feb 11;15(3):1311–1326. doi: 10.1093/nar/15.3.1311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dostatni N., Thierry F., Yaniv M. A dimer of BPV-1 E2 containing a protease resistant core interacts with its DNA target. EMBO J. 1988 Dec 1;7(12):3807–3816. doi: 10.1002/j.1460-2075.1988.tb03265.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Harrison S. M., Gearing K. L., Kim S. Y., Kingsman A. J., Kingsman S. M. Multiple cis-active elements in the long control region of bovine papillomavirus type 1 (BPV-1). Nucleic Acids Res. 1987 Dec 23;15(24):10267–10284. doi: 10.1093/nar/15.24.10267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Haugen T. H., Cripe T. P., Ginder G. D., Karin M., Turek L. P. Trans-activation of an upstream early gene promoter of bovine papilloma virus-1 by a product of the viral E2 gene. EMBO J. 1987 Jan;6(1):145–152. doi: 10.1002/j.1460-2075.1987.tb04732.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Haugen T. H., Turek L. P., Mercurio F. M., Cripe T. P., Olson B. J., Anderson R. D., Seidl D., Karin M., Schiller J. Sequence-specific and general transcriptional activation by the bovine papillomavirus-1 E2 trans-activator require an N-terminal amphipathic helix-containing E2 domain. EMBO J. 1988 Dec 20;7(13):4245–4253. doi: 10.1002/j.1460-2075.1988.tb03322.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Hermonat P. L., Spalholz B. A., Howley P. M. The bovine papillomavirus P2443 promoter is E2 trans-responsive: evidence for E2 autoregulation. EMBO J. 1988 Sep;7(9):2815–2822. doi: 10.1002/j.1460-2075.1988.tb03137.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Knight J. D., Li R., Botchan M. The activation domain of the bovine papillomavirus E2 protein mediates association of DNA-bound dimers to form DNA loops. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3204–3208. doi: 10.1073/pnas.88.8.3204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kost T. A., Theodorakis N., Hughes S. H. The nucleotide sequence of the chick cytoplasmic beta-actin gene. Nucleic Acids Res. 1983 Dec 10;11(23):8287–8301. doi: 10.1093/nar/11.23.8287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Lambert P. F., Spalholz B. A., Howley P. M. A transcriptional repressor encoded by BPV-1 shares a common carboxy-terminal domain with the E2 transactivator. Cell. 1987 Jul 3;50(1):69–78. doi: 10.1016/0092-8674(87)90663-5. [DOI] [PubMed] [Google Scholar]
  19. Li R., Knight J., Bream G., Stenlund A., Botchan M. Specific recognition nucleotides and their DNA context determine the affinity of E2 protein for 17 binding sites in the BPV-1 genome. Genes Dev. 1989 Apr;3(4):510–526. doi: 10.1101/gad.3.4.510. [DOI] [PubMed] [Google Scholar]
  20. McBride A. A., Byrne J. C., Howley P. M. E2 polypeptides encoded by bovine papillomavirus type 1 form dimers through the common carboxyl-terminal domain: transactivation is mediated by the conserved amino-terminal domain. Proc Natl Acad Sci U S A. 1989 Jan;86(2):510–514. doi: 10.1073/pnas.86.2.510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. McBride A. A., Schlegel R., Howley P. M. The carboxy-terminal domain shared by the bovine papillomavirus E2 transactivator and repressor proteins contains a specific DNA binding activity. EMBO J. 1988 Feb;7(2):533–539. doi: 10.1002/j.1460-2075.1988.tb02842.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Monini P., Grossman S. R., Pepinsky B., Androphy E. J., Laimins L. A. Cooperative binding of the E2 protein of bovine papillomavirus to adjacent E2-responsive sequences. J Virol. 1991 Apr;65(4):2124–2130. doi: 10.1128/jvi.65.4.2124-2130.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Morrissey L. C., Barsoum J., Androphy E. J. Trans activation by the bovine papillomavirus E2 protein in Saccharomyces cerevisiae. J Virol. 1989 Oct;63(10):4422–4425. doi: 10.1128/jvi.63.10.4422-4425.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Moskaluk C. A., Bastia D. Interaction of the bovine papillomavirus type 1 E2 transcriptional control protein with the viral enhancer: purification of the DNA-binding domain and analysis of its contact points with DNA. J Virol. 1988 Jun;62(6):1925–1931. doi: 10.1128/jvi.62.6.1925-1931.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Moskaluk C., Bastia D. The E2 "gene" of bovine papillomavirus encodes an enhancer-binding protein. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1215–1218. doi: 10.1073/pnas.84.5.1215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Prakash S. S., Grossman S. R., Pepinsky R. B., Laimins L. A., Androphy E. J. Amino acids necessary for DNA contact and dimerization imply novel motifs in the papillomavirus E2 trans-activator. Genes Dev. 1992 Jan;6(1):105–116. doi: 10.1101/gad.6.1.105. [DOI] [PubMed] [Google Scholar]
  27. Prakash S. S., Horwitz B. H., Zibello T., Settleman J., DiMaio D. Bovine papillomavirus E2 gene regulates expression of the viral E5 transforming gene. J Virol. 1988 Oct;62(10):3608–3613. doi: 10.1128/jvi.62.10.3608-3613.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Seiler-Tuyns A., Eldridge J. D., Paterson B. M. Expression and regulation of chicken actin genes introduced into mouse myogenic and nonmyogenic cells. Proc Natl Acad Sci U S A. 1984 May;81(10):2980–2984. doi: 10.1073/pnas.81.10.2980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sowden M., Harrison S., Ashfield R., Kingsman A. J., Kingsman S. M. Multiple cooperative interactions constrain BPV-1 E2 dependent activation of transcription. Nucleic Acids Res. 1989 Apr 25;17(8):2959–2972. doi: 10.1093/nar/17.8.2959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Spalholz B. A., Byrne J. C., Howley P. M. Evidence for cooperativity between E2 binding sites in E2 trans-regulation of bovine papillomavirus type 1. J Virol. 1988 Sep;62(9):3143–3150. doi: 10.1128/jvi.62.9.3143-3150.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Treacy M. N., He X., Rosenfeld M. G. I-POU: a POU-domain protein that inhibits neuron-specific gene activation. Nature. 1991 Apr 18;350(6319):577–584. doi: 10.1038/350577a0. [DOI] [PubMed] [Google Scholar]

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