Abstract
The noncoding region of the highly oncogenic, epidermodysplasia verruciformis-associated human papillomavirus type 8 contains a negative regulatory element (NRE). Quantitative RNase protection analysis confirmed that the NRE sequence acts as a silencer of transcription. A 38-bp sequence upstream of late promoter P7535 down-regulated expression from the homologous P7535 promoter, as well as the heterologous tk gene promoter, independently of its orientation relative to the test promoters. It also reduced gene expression when cloned downstream of the transcription units. Transient expression assays with keratinocytes and fibroblasts of epidermodysplasia verruciformis patients and controls demonstrated that the NRE activity is not cell specific. Gel retardation tests suggested that NRE specifically interacts with only one nuclear factor. Mutational analysis identified three NRE mutants which no longer formed a detectable DNA-protein complex but still repressed transcription, indicating that protein-DNA interaction is not relevant for the silencer function. The NRE contains a binding site of viral trans activator protein E2. It was shown that expression of E2 overrides the inhibitory effect of the NRE sequences. Binding of E2 and that of the cellular factor were mutually exclusive. The bifunctional nature of NRE acting as a silencer and a target site for viral trans activator E2 offers an interesting opportunity to regulate the switch from early to late transcription in the human papillomavirus life cycle.
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Selected References
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- Baniahmad A., Muller M., Steiner C., Renkawitz R. Activity of two different silencer elements of the chicken lysozyme gene can be compensated by enhancer elements. EMBO J. 1987 Aug;6(8):2297–2303. doi: 10.1002/j.1460-2075.1987.tb02504.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bartsch D., Boye B., Baust C., zur Hausen H., Schwarz E. Retinoic acid-mediated repression of human papillomavirus 18 transcription and different ligand regulation of the retinoic acid receptor beta gene in non-tumorigenic and tumorigenic HeLa hybrid cells. EMBO J. 1992 Jun;11(6):2283–2291. doi: 10.1002/j.1460-2075.1992.tb05287.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bauknecht T., Angel P., Royer H. D., zur Hausen H. Identification of a negative regulatory domain in the human papillomavirus type 18 promoter: interaction with the transcriptional repressor YY1. EMBO J. 1992 Dec;11(12):4607–4617. doi: 10.1002/j.1460-2075.1992.tb05563.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brand A. H., Breeden L., Abraham J., Sternglanz R., Nasmyth K. Characterization of a "silencer" in yeast: a DNA sequence with properties opposite to those of a transcriptional enhancer. Cell. 1985 May;41(1):41–48. doi: 10.1016/0092-8674(85)90059-5. [DOI] [PubMed] [Google Scholar]
- Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
- 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]
- Dürst M., Glitz D., Schneider A., zur Hausen H. Human papillomavirus type 16 (HPV 16) gene expression and DNA replication in cervical neoplasia: analysis by in situ hybridization. Virology. 1992 Jul;189(1):132–140. doi: 10.1016/0042-6822(92)90688-l. [DOI] [PubMed] [Google Scholar]
- Fogh J., Wright W. C., Loveless J. D. Absence of HeLa cell contamination in 169 cell lines derived from human tumors. J Natl Cancer Inst. 1977 Feb;58(2):209–214. doi: 10.1093/jnci/58.2.209. [DOI] [PubMed] [Google Scholar]
- Fuchs P. G., Iftner T., Weninger J., Pfister H. Epidermodysplasia verruciformis-associated human papillomavirus 8: genomic sequence and comparative analysis. J Virol. 1986 May;58(2):626–634. doi: 10.1128/jvi.58.2.626-634.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Furth P. A., Baker C. C. An element in the bovine papillomavirus late 3' untranslated region reduces polyadenylated cytoplasmic RNA levels. J Virol. 1991 Nov;65(11):5806–5812. doi: 10.1128/jvi.65.11.5806-5812.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gerlach K. K., Schloemer R. H. Hepatitis B virus C gene promoter is under negative regulation. Virology. 1992 Jul;189(1):59–66. doi: 10.1016/0042-6822(92)90681-e. [DOI] [PubMed] [Google Scholar]
- Gissmann L. Human papillomaviruses and genital cancer. Semin Cancer Biol. 1992 Oct;3(5):253–261. [PubMed] [Google Scholar]
- Goodbourn S. Negative regulation of transcriptional initiation in eukaryotes. Biochim Biophys Acta. 1990 Jun 1;1032(1):53–77. doi: 10.1016/0304-419x(90)90012-p. [DOI] [PubMed] [Google Scholar]
- 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]
- Higgins G. D., Uzelin D. M., Phillips G. E., McEvoy P., Marin R., Burrell C. J. Transcription patterns of human papillomavirus type 16 in genital intraepithelial neoplasia: evidence for promoter usage within the E7 open reading frame during epithelial differentiation. J Gen Virol. 1992 Aug;73(Pt 8):2047–2057. doi: 10.1099/0022-1317-73-8-2047. [DOI] [PubMed] [Google Scholar]
- Honda M., Niimura M. [Characterization of eccrine carcinoma established in tissue culture from a patient with epidermodysplasia verruciformis]. Nihon Hifuka Gakkai Zasshi. 1985 Feb;95(2):145–155. [PubMed] [Google Scholar]
- Horn S., Pfister H., Fuchs P. G. Constitutive transcriptional activator of Epidermodysplasia verruciformis-associated human papillomavirus 8. Virology. 1993 Oct;196(2):674–681. doi: 10.1006/viro.1993.1524. [DOI] [PubMed] [Google Scholar]
- Iftner T., Oft M., Böhm S., Wilczynski S. P., Pfister H. Transcription of the E6 and E7 genes of human papillomavirus type 6 in anogenital condylomata is restricted to undifferentiated cell layers of the epithelium. J Virol. 1992 Aug;66(8):4639–4646. doi: 10.1128/jvi.66.8.4639-4646.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jackson M. E., Campo M. S. Positive and negative E2-independent regulatory elements in the long control region of bovine papillomavirus type 4. J Gen Virol. 1991 Apr;72(Pt 4):877–883. doi: 10.1099/0022-1317-72-4-877. [DOI] [PubMed] [Google Scholar]
- Johnson P. F., McKnight S. L. Eukaryotic transcriptional regulatory proteins. Annu Rev Biochem. 1989;58:799–839. doi: 10.1146/annurev.bi.58.070189.004055. [DOI] [PubMed] [Google Scholar]
- Jones C., Zhu F., Dhanwada K. R. Analysis of a herpes simplex virus 2 fragment from the open reading frame of the large subunit of ribonucleotide reductase with transcriptional regulatory activity. DNA Cell Biol. 1993 Mar;12(2):127–137. doi: 10.1089/dna.1993.12.127. [DOI] [PubMed] [Google Scholar]
- Jones N. C., Rigby P. W., Ziff E. B. Trans-acting protein factors and the regulation of eukaryotic transcription: lessons from studies on DNA tumor viruses. Genes Dev. 1988 Mar;2(3):267–281. doi: 10.1101/gad.2.3.267. [DOI] [PubMed] [Google Scholar]
- Kennedy I. M., Haddow J. K., Clements J. B. A negative regulatory element in the human papillomavirus type 16 genome acts at the level of late mRNA stability. J Virol. 1991 Apr;65(4):2093–2097. doi: 10.1128/jvi.65.4.2093-2097.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kyo S., Inoue M., Nishio Y., Nakanishi K., Akira S., Inoue H., Yutsudo M., Tanizawa O., Hakura A. NF-IL6 represses early gene expression of human papillomavirus type 16 through binding to the noncoding region. J Virol. 1993 Feb;67(2):1058–1066. doi: 10.1128/jvi.67.2.1058-1066.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Levine M., Manley J. L. Transcriptional repression of eukaryotic promoters. Cell. 1989 Nov 3;59(3):405–408. doi: 10.1016/0092-8674(89)90024-x. [DOI] [PubMed] [Google Scholar]
- Luckow B., Schütz G. CAT constructions with multiple unique restriction sites for the functional analysis of eukaryotic promoters and regulatory elements. Nucleic Acids Res. 1987 Jul 10;15(13):5490–5490. doi: 10.1093/nar/15.13.5490. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maniatis T., Goodbourn S., Fischer J. A. Regulation of inducible and tissue-specific gene expression. Science. 1987 Jun 5;236(4806):1237–1245. doi: 10.1126/science.3296191. [DOI] [PubMed] [Google Scholar]
- May M., Dong X. P., Beyer-Finkler E., Stubenrauch F., Fuchs P. G., Pfister H. The E6/E7 promoter of extrachromosomal HPV16 DNA in cervical cancers escapes from cellular repression by mutation of target sequences for YY1. EMBO J. 1994 Mar 15;13(6):1460–1466. doi: 10.1002/j.1460-2075.1994.tb06400.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- May M., Helbl V., Pfister H., Fuchs P. G. Unique topography of DNA-protein interactions in the non-coding region of epidermodysplasia verruciformis-associated human papillomaviruses. J Gen Virol. 1991 Dec;72(Pt 12):2989–2997. doi: 10.1099/0022-1317-72-12-2989. [DOI] [PubMed] [Google Scholar]
- 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]
- Montalvo E. A., Shi Y., Shenk T. E., Levine A. J. Negative regulation of the BZLF1 promoter of Epstein-Barr virus. J Virol. 1991 Jul;65(7):3647–3655. doi: 10.1128/jvi.65.7.3647-3655.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nilsson T., Sjöblom A., Masucci M. G., Rymo L. Viral and cellular factors influence the activity of the Epstein-Barr virus BCR2 and BWR1 promoters in cells of different phenotype. Virology. 1993 Apr;193(2):774–785. doi: 10.1006/viro.1993.1186. [DOI] [PubMed] [Google Scholar]
- Paciucci R., Pellicer A. Dissection of the mouse N-ras gene upstream regulatory sequences and identification of the promoter and a negative regulatory element. Mol Cell Biol. 1991 Mar;11(3):1334–1343. doi: 10.1128/mcb.11.3.1334. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pfister H. Biology and biochemistry of papillomaviruses. Rev Physiol Biochem Pharmacol. 1984;99:111–181. doi: 10.1007/BFb0027716. [DOI] [PubMed] [Google Scholar]
- Pfister H. Human papillomaviruses and skin cancer. Semin Cancer Biol. 1992 Oct;3(5):263–271. [PubMed] [Google Scholar]
- Pierce A. J., Jambou R. C., Jensen D. E., Azizkhan J. C. A conserved DNA structural control element modulates transcription of a mammalian gene. Nucleic Acids Res. 1992 Dec 25;20(24):6583–6587. doi: 10.1093/nar/20.24.6583. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ptashne M., Gann A. A. Activators and targets. Nature. 1990 Jul 26;346(6282):329–331. doi: 10.1038/346329a0. [DOI] [PubMed] [Google Scholar]
- Reh H., Pfister H. Human papillomavirus type 8 contains cis-active positive and negative transcriptional control sequences. J Gen Virol. 1990 Oct;71(Pt 10):2457–2462. doi: 10.1099/0022-1317-71-10-2457. [DOI] [PubMed] [Google Scholar]
- Renkawitz R. Transcriptional repression in eukaryotes. Trends Genet. 1990 Jun;6(6):192–197. doi: 10.1016/0168-9525(90)90176-7. [DOI] [PubMed] [Google Scholar]
- 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]
- Schiffman M. H. Recent progress in defining the epidemiology of human papillomavirus infection and cervical neoplasia. J Natl Cancer Inst. 1992 Mar 18;84(6):394–398. doi: 10.1093/jnci/84.6.394. [DOI] [PubMed] [Google Scholar]
- Sealey L., Chalkley R. At least two nuclear proteins bind specifically to the Rous sarcoma virus long terminal repeat enhancer. Mol Cell Biol. 1987 Feb;7(2):787–798. doi: 10.1128/mcb.7.2.787. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stoler M. H., Rhodes C. R., Whitbeck A., Wolinsky S. M., Chow L. T., Broker T. R. Human papillomavirus type 16 and 18 gene expression in cervical neoplasias. Hum Pathol. 1992 Feb;23(2):117–128. doi: 10.1016/0046-8177(92)90232-r. [DOI] [PubMed] [Google Scholar]
- Stubenrauch F., Malejczyk J., Fuchs P. G., Pfister H. Late promoter of human papillomavirus type 8 and its regulation. J Virol. 1992 Jun;66(6):3485–3493. doi: 10.1128/jvi.66.6.3485-3493.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sutcliffe J. G. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. doi: 10.1101/sqb.1979.043.01.013. [DOI] [PubMed] [Google Scholar]
- Westin G., Gerster T., Müller M. M., Schaffner G., Schaffner W. OVEC, a versatile system to study transcription in mammalian cells and cell-free extracts. Nucleic Acids Res. 1987 Sep 11;15(17):6787–6798. doi: 10.1093/nar/15.17.6787. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wu T. C., Mounts P. Transcriptional regulatory elements in the noncoding region of human papillomavirus type 6. J Virol. 1988 Dec;62(12):4722–4729. doi: 10.1128/jvi.62.12.4722-4729.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yasumoto S., Taniguchi A., Sohma K. Epidermal growth factor (EGF) elicits down-regulation of human papillomavirus type 16 (HPV-16) E6/E7 mRNA at the transcriptional level in an EGF-stimulated human keratinocyte cell line: functional role of EGF-responsive silencer in the HPV-16 long control region. J Virol. 1991 Apr;65(4):2000–2009. doi: 10.1128/jvi.65.4.2000-2009.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de Wet J. R., Wood K. V., DeLuca M., Helinski D. R., Subramani S. Firefly luciferase gene: structure and expression in mammalian cells. Mol Cell Biol. 1987 Feb;7(2):725–737. doi: 10.1128/mcb.7.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]