Prebending the estrogen response element destabilizes binding of the estrogen receptor DNA binding domain

Jongsook Kim, Georgius De Haan, Ann M. Nardulli, David J. Shapiro

Research output: Contribution to journalArticlepeer-review

Abstract

Binding of many eukaryotic transcription regulatory proteins to their DNA recognition sequences results in conformational changes in DNA. To test the effect of altering DNA topology by prebending a transcription factor binding site, we examined the interaction of the estrogen receptor (ER) DNA binding domain (DBD) with prebent estrogen response elements (EREs). When the ERE in minicircle DNA was prebent toward the major groove, which is in the same direction as the ER-induced DNA bend, there was no significant effect on ER DBD binding relative to the linear counterparts. However, when the ERE was bent toward the minor groove, in a direction that opposes the ER-induced DNA bend, there was a four- tn eightfold reduction in ER DBD binding. Since reduced binding was also observed with the ERE in nicked circles, the reduction in binding was not due to torsional force induced by binding of ER DBD to the prebent ERE in covalently closed minicircles. To determine the mechanism responsible for reduced binding to the prebent ERE, we examined the effect of prebending the ERE on the association and dissociation of the ER DBD. Binding of the ER DBD to ERE-containing minicircles was rapid when the EREs were prebent toward either the major or minor groove of the DNA (k(on) of 9.9.106 to 1.7. 107M-1 s-1). Prebending the ERE toward the minor groove resulted in an increase in k(of) of four- to fivefold. Increased dissociation of the ER DBD from the ERE is, therefore, the major factor responsible for reduced binding of the ER DBD to an ERE prebent toward the minor groove. These data provide the first direct demonstration that the interaction of a eukaryotic transcription factor with its recognition sequence can be strongly influenced by altering DNA topology through prebending the DNA.

Original languageEnglish (US)
Pages (from-to)3173-3180
Number of pages8
JournalMolecular and cellular biology
Volume17
Issue number6
DOIs
StatePublished - Jun 1997

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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