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J. Biol. Chem., Vol. 276, Issue 51, 48562-48571, December 21, 2001
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From the University of Wisconsin Medical School,
Madison, Wisconsin 53706
The c-Myc protein is a site-specific
DNA-binding transcription factor that is up-regulated in a number of
different cancers. We have previously shown that binding of Myc
correlates with increased transcription of the cad
promoter. We have now further investigated the mechanism by which Myc
mediates transcriptional activation of the cad gene.
Using a chromatin immunoprecipitation assay, we found high levels of
RNA polymerase II bound to the cad promoter in quiescent
NIH 3T3 cells and in differentiated U937 cells, even though the
promoter is inactive. However, chromatin immunoprecipitation with an
antibody that recognizes the hyperphosphorylated form of the RNA
polymerase II carboxyl-terminal domain (CTD) revealed that
phosphorylation of the CTD does correlate with c-Myc binding and
cad transcription. We have also found that the c-Myc
transactivation domain interacts with cdk9 and cyclin T1, components of
the CTD kinase P-TEFb. Furthermore, activator bypass experiments have shown that direct recruitment of cyclin T1 to the cad
promoter can substitute for c-Myc to activate the promoter. In summary, our results suggest that c-Myc activates transcription of
cad by stimulating promoter clearance and elongation,
perhaps via recruitment of P-TEFb.
c-Myc Mediates Activation of the cad Promoter via a
Post-RNA Polymerase II Recruitment Mechanism*
*
This work was supported in part by United States Public
Health Service Grants CA07175, CA09135, and CA45240 from the National Institutes of Health.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: McArdle Laboratory for
Cancer Research, University of Wisconsin Medical School, Madison, WI
53706. Tel.: 608-262-2071; Fax: 608-262-2824; E-mail: farnham@oncology.wisc.edu.
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