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Papers In Press, published online ahead of print August 24, 2001
J. Biol. Chem, 10.1074/jbc.M103615200
Submitted on April 23, 2001
Revised on August 22, 2001
Accepted on August 24, 2001
Pharmacological and Physiological Sciences, Saint Louis University School of Medicine, Saint Louis, MI 63104
Corresponding Author: CHRIVIA{at}slu.edu
SRCAP belongs to the SNF2 family of proteins whose members participate in various aspects of transcriptional regulation, including chromatin remodeling. It was identified by its ability to bind to CREB-Binding Protein (CBP) and it increases the transactivation function of CBP. The PEPCK promoter was used as a model system to explore the role of SRCAP in the regulation of transcription mediated by factors which utilize CBP as a coactivator. We show that transcription of a PEPCK CAT reporter gene activated by PKA, is enhanced 7-fold by SRCAP. In the absence of PKA this SRCAP-mediated enhancement does not occur, suggesting SRCAP functions as a coactivator for PKA activated factors such as CREB. Replacing the PEPCK promoter binding site for CREB with a binding site for Gal 4 (*CRE Gal4 PEPCK-CAT reporter gene) blocks the ability of SRCAP to activate transcription despite the presence of PKA. Expression of a Gal-CREB chimera restores the ability of PKA to regulate transcription of the * CRE Gal4 PEPCK gene and restored the ability of SRCAP to stimulate PKA-activated transcription. In addition, SRCAP in the presence of PKA enhances the ability of the Gal-CREB chimera to activate transcription of a Gal-CAT reporter gene which contains only binding sites for Gal 4. SRCAP binds to CBP amino acids 280-460, a region that is important for CBP to function as a coactivator for CREB. Overexpression of a SRCAP peptide corresponding to this CBP binding domain acts as a dominant negative inhibitor of CREB-mediated transcription. Structure-function studies were done to explore the mechanism(s) by which SRCAP regulates transcription. These studies indicate that the N-terminal region of SRCAP, which contains five of the seven regions that comprise the ATPase domain, is not needed for activation of CREB-mediated transcription. SRCAP apparently has several domains that participate in the activation of transcription.
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