Electrostatic control of half-site spacing preferences by the cyclic AMP response element-binding protein CREB Export

@article{citeulike:2364022, abstract = {Basic region leucine zipper (bZIP) proteins represent a class of transcription factors that bind DNA using a simple, dimeric, {alpha}-helical recognition motif. The cAMP response element-binding protein (CREB) is a member of the CREB/ATF subfamily of bZIP proteins. CREB discriminates effectively in vivo and in vitro between the 10 bp cAMP response element (ATGACGTCAT, CRE) and the 9 bp activating protein 1 site (ATGACTCAT, AP-1). Here we describe an alanine scanning mutagenesis study designed to identify those residues within the CREB bZIP element that control CRE/AP-1 specificity. We find that the preference of CREB for the CRE site is controlled in a positive and negative way by acidic and basic residues in the basic, spacer and zipper segments. The CRE/AP-1 specificity of CREB is increased significantly by four glutamic acid residues located at positions 24, 28, 35 and 41; glutamic acid residues at positions 10 and 48 contribute in a more modest way. Specificity is decreased significantly by two basic residues located at positions 21 and 23; basic residues at positions 14, 18, 33 and 34 and V17 contribute in a more modest way. All of the residues that influence specificity significantly are located on the solvent-exposed face of the protein-DNA complex and likely participate in interactions between and among proteins, not between protein and DNA. The finding that the CRE/AP-1 specificity of CREB is dictated by the presence or absence of charged residues has interesting implications for how transcription factors seek and selectively bind sequences within genomic DNA. 10.1093/nar/29.16.3311}, author = {Montclare, Jin K. and Sloan, Leslie S. and Schepartz, Alanna}, citeulike-article-id = {2364022}, citeulike-linkout-0 = {http://dx.doi.org/10.1093/nar/29.16.3311}, citeulike-linkout-1 = {http://nar.oxfordjournals.org/cgi/content/abstract/29/16/3311}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/11504868}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=11504868}, day = {15}, doi = {10.1093/nar/29.16.3311}, journal = {Nucl. Acids Res.}, keywords = {binding\_specificity, bzip}, month = {August}, number = {16}, pages = {3311--3319}, posted-at = {2008-02-11 22:27:04}, priority = {2}, title = {Electrostatic control of half-site spacing preferences by the cyclic AMP response element-binding protein CREB}, url = {http://dx.doi.org/10.1093/nar/29.16.3311}, volume = {29}, year = {2001} }

TY - JOUR ID - citeulike:2364022 L3 - citeulike-article-id:2364022 N2 - Basic region leucine zipper (bZIP) proteins represent a class of transcription factors that bind DNA using a simple, dimeric, {alpha}-helical recognition motif. The cAMP response element-binding protein (CREB) is a member of the CREB/ATF subfamily of bZIP proteins. CREB discriminates effectively in vivo and in vitro between the 10 bp cAMP response element (ATGACGTCAT, CRE) and the 9 bp activating protein 1 site (ATGACTCAT, AP-1). Here we describe an alanine scanning mutagenesis study designed to identify those residues within the CREB bZIP element that control CRE/AP-1 specificity. We find that the preference of CREB for the CRE site is controlled in a positive and negative way by acidic and basic residues in the basic, spacer and zipper segments. The CRE/AP-1 specificity of CREB is increased significantly by four glutamic acid residues located at positions 24, 28, 35 and 41; glutamic acid residues at positions 10 and 48 contribute in a more modest way. Specificity is decreased significantly by two basic residues located at positions 21 and 23; basic residues at positions 14, 18, 33 and 34 and V17 contribute in a more modest way. All of the residues that influence specificity significantly are located on the solvent-exposed face of the protein-DNA complex and likely participate in interactions between and among proteins, not between protein and DNA. The finding that the CRE/AP-1 specificity of CREB is dictated by the presence or absence of charged residues has interesting implications for how transcription factors seek and selectively bind sequences within genomic DNA. 10.1093/nar/29.16.3311 IS - 16 JF - Nucl. Acids Res. EP - 3319 TI - Electrostatic control of half-site spacing preferences by the cyclic AMP response element-binding protein CREB VL - 29 SP - 3311 KW - binding_specificity KW - bzip AU - Montclare, Jin AU - Sloan, Leslie AU - Schepartz, Alanna PY - 2001/08/15/ UR - http://dx.doi.org/10.1093/nar/29.16.3311 ER -