This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Stünkel, W.
Right arrow Articles by Bernard, H.-U.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Stünkel, W.
Right arrow Articles by Bernard, H.-U.

Next Article 

Journal of Virology, March 2000, p. 2489-2501, Vol. 74, No. 6
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Nuclear Matrix Attachment Regions of Human Papillomavirus Type 16 Repress or Activate the E6 Promoter, Depending on the Physical State of the Viral DNA

Walter Stünkel, Zhonghui Huang, Shyh-Han Tan,dagger Mark J. O'Connor,Dagger and Hans-Ulrich Bernard*

Institute of Molecular and Cell Biology, National University of Singapore, Singapore 117609, Republic of Singapore

Received 1 October 1999/Accepted 15 December 1999

Two nuclear matrix attachment regions (MARs) bracket a 550-bp segment of the long control region (LCR) containing the epithelial cell-specific enhancer and the E6 promoter of human papillomavirus type 16 (HPV-16). One of these MARs is located in the 5' third of the LCR (5'-LCR-MAR); the other lies within the E6 gene (E6-MAR). To study their function, we linked these MARs in various natural or artificial permutations to a chimeric gene consisting of the HPV-16 enhancer-promoter segment and a reporter gene. In transient transfections of HeLa cells, the presence of either of these two MARs strongly represses reporter gene expression. In contrast to this, but similar to the published behavior of cellular MARs, reporter gene expression is stimulated strongly by the E6-MAR and moderately by the 5'-LCR-MAR in stable transfectants of HeLa or C33A cells. To search for binding sites of soluble nuclear proteins which may be responsible for repression during transient transfections, we performed electrophoretic mobility shift assays (EMSAs) of overlapping oligonucleotides that represented all sequences of these two MARs. Both MARs contain multiple sites for two strongly binding proteins and weak binding sites for additional factors. The strongest complex, with at least five binding sites in each MAR, is generated by the CCAAT displacement factor (CDP)/Cut, as judged by biochemical purification, by EMSAs with competing oligonucleotides and with anti-CDP/Cut oligonucleotides, and by mutations. CDP/Cut, a repressor that is down-regulated during differentiation, apparently represses HPV-16 transcription in undifferentiated epithelials cells and in HeLa cells, which are rich in CDP/Cut. In analogy to poorly understood mechanisms acting on cellular MARs, activation after physical linkage to chromosomal DNA may result from competition between the nuclear matrix and CDP/Cut. Our observations show that cis-responsive elements that regulate the HPV-16 E6 promoter are tightly clustered over at least 1.3 kb and occur throughout the E6 gene. HPV-16 MARs are context dependent transcriptional enhancers, and activated expression of HPV-16 oncogenes dependent on chromosomal integration may positively select tumorigenic cells during the multistep etiology of cervical cancer.

* Corresponding author. Mailing address: Institute of Molecular and Cell Biology, National University of Singapore, 30 Medical Dr., Singapore 117609, Republic of Singapore. Phone: 65-778-8823. Fax: 65-779-1117. E-mail: mcbhub{at}imcb.nus.edu.sg.

dagger Present address: National Institutes of Health, Unit of Cell Cycle Regulation, Bethesda, MD 20892-5431.

Dagger Present address: KuDOS Pharmaceuticals Ltd., Cambridge CB4 4GW, United Kingdom.

Journal of Virology, March 2000, p. 2489-2501, Vol. 74, No. 6
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Truscott, M., Harada, R., Vadnais, C., Robert, F., Nepveu, A. (2008). p110 CUX1 Cooperates with E2F Transcription Factors in the Transcriptional Activation of Cell Cycle-Regulated Genes. Mol. Cell. Biol. 28: 3127-3138 [Abstract] [Full Text]  
  • Harada, R., Vadnais, C., Sansregret, L., Leduy, L., Berube, G., Robert, F., Nepveu, A. (2008). Genome-wide location analysis and expression studies reveal a role for p110 CUX1 in the activation of DNA replication genes. Nucleic Acids Res 36: 189-202 [Abstract] [Full Text]  
  • Sansregret, L., Goulet, B., Harada, R., Wilson, B., Leduy, L., Bertoglio, J., Nepveu, A. (2006). The p110 Isoform of the CDP/Cux Transcription Factor Accelerates Entry into S Phase.. Mol. Cell. Biol. 26: 2441-2455 [Abstract] [Full Text]  
  • Kalantari, M., Calleja-Macias, I. E., Tewari, D., Hagmar, B., Lie, K., Barrera-Saldana, H. A., Wiley, D. J., Bernard, H.-U. (2004). Conserved Methylation Patterns of Human Papillomavirus Type 16 DNA in Asymptomatic Infection and Cervical Neoplasia. J. Virol. 78: 12762-12772 [Abstract] [Full Text]  
  • Zimmer, C., Tiveron, M.-C., Bodmer, R., Cremer, H. (2004). Dynamics of Cux2 Expression Suggests that an Early Pool of SVZ Precursors is Fated to Become Upper Cortical Layer Neurons. Cereb Cortex 14: 1408-1420 [Abstract] [Full Text]  
  • Truscott, M., Raynal, L., Wang, Y., Berube, G., Leduy, L., Nepveu, A. (2004). The N-terminal Region of the CCAAT Displacement Protein (CDP)/Cux Transcription Factor Functions as an Autoinhibitory Domain that Modulates DNA Binding. J. Biol. Chem. 279: 49787-49794 [Abstract] [Full Text]  
  • Kaul-Ghanekar, R., Jalota, A., Pavithra, L., Tucker, P., Chattopadhyay, S. (2004). SMAR1 and Cux/CDP modulate chromatin and act as negative regulators of the TCR{beta} enhancer (E{beta}). Nucleic Acids Res 32: 4862-4875 [Abstract] [Full Text]  
  • Rudd, S., Frisch, M., Grote, K., Meyers, B. C., Mayer, K., Werner, T. (2004). Genome-Wide in Silico Mapping of Scaffold/Matrix Attachment Regions in Arabidopsis Suggests Correlation of Intragenic Scaffold/Matrix Attachment Regions with Gene Expression. Plant Physiol. 135: 715-722 [Abstract] [Full Text]  
  • Mearini, G., Nielsen, P. E., Fackelmayer, F. O. (2004). Localization and dynamics of small circular DNA in live mammalian nuclei. Nucleic Acids Res 32: 2642-2651 [Abstract] [Full Text]  
  • Glahder, J. A., Hansen, C. N., Vinther, J., Madsen, B. S., Norrild, B. (2003). A promoter within the E6 ORF of human papillomavirus type 16 contributes to the expression of the E7 oncoprotein from a monocistronic mRNA. J. Gen. Virol. 84: 3429-3441 [Abstract] [Full Text]  
  • Kim, K., Garner-Hamrick, P. A., Fisher, C., Lee, D., Lambert, P. F. (2003). Methylation Patterns of Papillomavirus DNA, Its Influence on E2 Function, and Implications in Viral Infection. J. Virol. 77: 12450-12459 [Abstract] [Full Text]  
  • Rosenstierne, M. W., Vinther, J., Hansen, C. N., Prydsoe, M., Norrild, B. (2003). Identification and characterization of a cluster of transcription start sites located in the E6 ORF of human papillomavirus type 16. J. Gen. Virol. 84: 2909-2920 [Abstract] [Full Text]  
  • Fujii, T., Austin, D., Guo, D., Srimatkandada, S., Wang, T., Kubushiro, K., Masumoto, N., Tsukazaki, K., Nozawa, S., Deisseroth, A. B. (2003). Peptides Inhibitory for the Transcriptional Regulatory Function of Human Papillomavirus E2. Clin. Cancer Res. 9: 5423-5428 [Abstract] [Full Text]  
  • Badal, V., Chuang, L. S. H., Tan, E. H.-H., Badal, S., Villa, L. L., Wheeler, C. M., Li, B. F. L., Bernard, H.-U. (2003). CpG Methylation of Human Papillomavirus Type 16 DNA in Cervical Cancer Cell Lines and in Clinical Specimens: Genomic Hypomethylation Correlates with Carcinogenic Progression. J. Virol. 77: 6227-6234 [Abstract] [Full Text]  
  • Truscott, M., Raynal, L., Premdas, P., Goulet, B., Leduy, L., Berube, G., Nepveu, A. (2003). CDP/Cux Stimulates Transcription from the DNA Polymerase {alpha} Gene Promoter. Mol. Cell. Biol. 23: 3013-3028 [Abstract] [Full Text]  
  • Lukaszuk, K., Liss, J., Wozniak, I., Emerich, J., Wojcikowski, C. (2003). Human Papillomavirus Type 16 Status in Cervical Carcinoma Cell DNA Assayed by Multiplex PCR. J. Clin. Microbiol. 41: 608-612 [Abstract] [Full Text]  
  • Goulet, B., Watson, P., Poirier, M., Leduy, L., Berube, G., Meterissian, S., Jolicoeur, P., Nepveu, A. (2002). Characterization of a Tissue-specific CDP/Cux Isoform, p75, Activated in Breast Tumor Cells. Cancer Res. 62: 6625-6633 [Abstract] [Full Text]  
  • Santaguida, M., Ding, Q., Berube, G., Truscott, M., Whyte, P., Nepveu, A. (2001). Phosphorylation of the CCAAT Displacement Protein (CDP)/Cux Transcription Factor by Cyclin A-Cdk1 Modulates Its DNA Binding Activity in G2. J. Biol. Chem. 276: 45780-45790 [Abstract] [Full Text]  
  • Vance, K. W., Campo, M. S., Morgan, I. M. (2001). A Novel Silencer Element in the Bovine Papillomavirus Type 4 Promoter Represses the Transcriptional Response to Papillomavirus E2 Protein. J. Virol. 75: 2829-2838 [Abstract] [Full Text]  
  • Antes, T. J., Chen, J., Cooper, A. D., Levy-Wilson, B. (2000). The Nuclear Matrix Protein CDP Represses Hepatic Transcription of the Human Cholesterol-7alpha Hydroxylase Gene. J. Biol. Chem. 275: 26649-26660 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»