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M411147200v1
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Papers In Press, published online ahead of print November 19, 2004
J. Biol. Chem, 10.1074/jbc.M411147200
Submitted on September 29, 2004
Revised on November 10, 2004
Accepted on November 19, 2004

Chromatin-dependent E1A activity modulates NF-B Rel A-mediated repression of glucocorticoid receptor-dependent transcription

Barbara A. Burkhart, Pratibha B. Hebbar, Kevin W. Trotter, and Trevor K. Archer

Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709

Corresponding Author: archer1{at}niehs.nih.gov

The role of chromatin-dependent regulatory mechanisms in repression of glucocorticoid-dependent transcription from the MMTV promoter by p65 and E1A was investigated using chromatin and transiently transfected reporters. The p65 RelA subunit of NF-kB represses MMTV expression on either transient or an integrated reporters. In contrast, the viral oncoprotein E1A represses a transient but not an integrated MMTV. E1A repression is attenuated by chromatin, suggesting p65 but not E1A, manipulates chromatin appropriately to inhibit the GR. Co-expression of p65 and E1A additively represses the transient MMTV but restores the transcriptional activation of the chromatin MMTV in response to glucocorticoids. This indicates E1A has a dominant chromatin-dependent activity that attenuates repression by p65. E1A, p65 and GR bind the MMTV promoter and chromatin remodeling enhances binding on both repressed and activated promoters. In addition repression by p65 requires Brg for repression of the integrated MMTV. This suggests that neither p65 repression nor E1A attenuation of repression results from an inhibition of remodeling that prevents transcription factor binding. Further, p300/CBP is also required for both repression and attenuation by p65 and E1A since p65 and E1A p300/CBP-binding mutants are inactive, indicative of a requirement for p300/CBP-dependent complex formation for both repression and attenuation with chromatin. These data suggest that both the p65 dependent repression and the E1A mediated attenuation of repression require the Brg1-dependent chromatin remodeling function and p300/CBP-dependent complex formation at a promoter assembled within chromatin


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