This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplemental material
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 Hearnes, J. M.
Right arrow Articles by Pietenpol, J. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hearnes, J. M.
Right arrow Articles by Pietenpol, J. A.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, November 2005, p. 10148-10158, Vol. 25, No. 22
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.22.10148-10158.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Chromatin Immunoprecipitation-Based Screen To Identify Functional Genomic Binding Sites for Sequence-Specific Transactivators{dagger}

Jamie M. Hearnes, Deborah J. Mays, Kristy L. Schavolt, Luojia Tang, Xin Jiang, and Jennifer A. Pietenpol*

Department of Biochemistry, Center for Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, Tennessee 37232

Received 13 April 2005/ Returned for modification 6 June 2005/ Accepted 28 August 2005

In various human diseases, altered gene expression patterns are often the result of deregulated gene-specific transcription factor activity. To further understand disease on a molecular basis, the comprehensive analysis of transcription factor signaling networks is required. We developed an experimental approach, combining chromatin immunoprecipitation (ChIP) with a yeast-based assay, to screen the genome for transcription factor binding sites that link to transcriptionally regulated target genes. We used the tumor suppressor p53 to demonstrate the effectiveness of the method. Using primary and immortalized, nontransformed cultures of human mammary epithelial cells, we isolated over 100 genomic DNA fragments that contain novel p53 binding sites. This approach led to the identification and validation of novel p53 target genes involved in diverse signaling pathways, including growth factor signaling, protein kinase/phosphatase signaling, and RNA binding. Our results yield a more complete understanding of p53-regulated signaling pathways, and this approach could be applied to any number of transcription factors to further elucidate complex transcriptional networks.

* Corresponding author. Mailing address: 652 Preston Research Building, Vanderbilt-Ingram Cancer Center, Nashville, TN 37232-6838. Phone: (615) 936-1512. Fax: (615) 936-1790. E-mail: j.pietenpol{at}vanderbilt.edu.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

Molecular and Cellular Biology, November 2005, p. 10148-10158, Vol. 25, No. 22
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.22.10148-10158.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Brazdova, M., Quante, T., Togel, L., Walter, K., Loscher, C., Tichy, V., Cincarova, L., Deppert, W., Tolstonog, G. V. (2009). Modulation of gene expression in U251 glioblastoma cells by binding of mutant p53 R273H to intronic and intergenic sequences. Nucleic Acids Res 37: 1486-1500 [Abstract] [Full Text]  
  • Neilsen, P. M., Cheney, K. M., Li, C.-W., Chen, J. D., Cawrse, J. E., Schulz, R. B., Powell, J. A., Kumar, R., Callen, D. F. (2008). Identification of ANKRD11 as a p53 coactivator. J. Cell Sci. 121: 3541-3552 [Abstract] [Full Text]  
  • Rosenbluth, J. M., Mays, D. J., Pino, M. F., Tang, L. J., Pietenpol, J. A. (2008). A Gene Signature-Based Approach Identifies mTOR as a Regulator of p73. Mol. Cell. Biol. 28: 5951-5964 [Abstract] [Full Text]  
  • Ogden, S. R., Wroblewski, L. E., Weydig, C., Romero-Gallo, J., O'Brien, D. P., Israel, D. A., Krishna, U. S., Fingleton, B., Reynolds, A. B., Wessler, S., Peek, R. M. Jr (2008). p120 and Kaiso Regulate Helicobacter pylori-induced Expression of Matrix Metalloproteinase-7. Mol. Biol. Cell 19: 4110-4121 [Abstract] [Full Text]  
  • Smeenk, L., van Heeringen, S. J., Koeppel, M., van Driel, M. A., Bartels, S. J. J., Akkers, R. C., Denissov, S., Stunnenberg, H. G., Lohrum, M. (2008). Characterization of genome-wide p53-binding sites upon stress response. Nucleic Acids Res 36: 3639-3654 [Abstract] [Full Text]  
  • Menendez, D., Inga, A., Snipe, J., Krysiak, O., Schonfelder, G., Resnick, M. A. (2007). A Single-Nucleotide Polymorphism in a Half-Binding Site Creates p53 and Estrogen Receptor Control of Vascular Endothelial Growth Factor Receptor 1. Mol. Cell. Biol. 27: 2590-2600 [Abstract] [Full Text]  
  • Krieg, A. J., Hammond, E. M., Giaccia, A. J. (2006). Functional Analysis of p53 Binding under Differential Stresses.. Mol. Cell. Biol. 26: 7030-7045 [Abstract] [Full Text]  
  • Jacobs, F. M. J., Smits, S. M., Hornman, K. J. M., Burbach, J. P. H., Smidt, M. P. (2006). Strategies to unravel molecular codes essential for the development of meso-diencephalic dopaminergic neurons. J. Physiol. 575: 397-402 [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»