Consensus and Variant cAMP-regulated Enhancers Have Distinct CREB-binding Properties

Johanna C. Craig, Maria A. Schumacher, Steven E. Mansoor, David L. Farrens, Richard G. Brennan, Richard H. Goodman

Research output: Contribution to journalArticlepeer-review

61 Scopus citations


Recent determination of the cAMP response element-binding protein (CREB) basic leucine zipper (bZIP) consensus CRE crystal structure revealed key dimerization and DNA binding features that are conserved among members of the CREB/CREM/ATF-1 family of transcription factors. Dimerization appeared to be mediated by a Tyr307-Glus312 interhelical hydrogen bond and a Glu319-Arg314 electrostatic interaction. An unexpected hexahydrated Mg2+ ion was centered above the CRE in the dimer cavity. In the present study, we related these features to CREB dimerization and DNA binding. A Y307F substitution reduced dimer stability and DNA binding affinity, whereas a Y307R mutation produced a stabilizing effect. Mutation of Glu319 to Ala or Lys attenuated dimerization and DNA binding. Mg2+ ions enhanced the binding affinity of wild-type CREB to the palindromic CRE by ∼20-fold but did not do so for divergent CREs. Similarly, mutation of Lys304, which mediates the CREB interaction with the hydrated Mg2+, blocked CREB binding to the palindromic but not the variant CRE sequences. The distinct binding characteristics of the K304A mutants to the consensus and variant CRE sequences indicate that CREB binding to these elements is differentially regulated by Mg2+ ions. We suggest that CREB binds the consensus and variant CRE sequences through fundamentally distinct mechanisms.

Original languageEnglish (US)
Pages (from-to)11719-11728
Number of pages10
JournalJournal of Biological Chemistry
Issue number15
StatePublished - Apr 13 2001

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


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