Indirect readout of DNA sequence at the primary-kink site in the CAP-DNA complex: DNA binding specificity based on energetics of dna kinking

Shengfeng Chen, Angelo Gunasekera, Xiaoping Zhang, Thomas A. Kunkel, Richard H. Ebright, Helen M. Berman

Research output: Contribution to journalArticlepeer-review

47 Scopus citations

Abstract

The catabolite activator protein (CAP) makes no direct contact with the consensus base-pair T:A at position 6 of the DNA half-site 5′-A1A2A3T4G5T 6G7A8T9C10T 11-3′ but, nevertheless, exhibits strong specificity for T:A at position 6. Binding of CAP results in formation of a sharp DNA kink, with a roll angle of ∼40° and a twist angle of ∼20°, between positions 6 and 7 of the DNA half-site. The consensus base-pair T:A at position 6 and the consensus base-pair G:C at position 7 form a T:A/G:C step, which is known to be associated with DNA flexibility. It has been proposed that specificity for T:A at position 6 is a consequence of formation of the DNA kink between positions 6 and 7, and of effects of the T:A6/G:C7 step on the geometry of DNA kinking, or the energetics of DNA kinking. In this work, we determine crystallographic structures of CAP-DNA complexes having the consensus base-pair T:A at position 6 or the non-consensus base-pair C:G at position 6. We show that complexes containing T:A or C:G at position 6 exhibit similar overall DNA bend angles and local geometries of DNA kinking. We infer that indirect readout in this system does not involve differences in the geometry of DNA kinking but, rather, solely differences in the energetics of DNA kinking. We further infer that the main determinant of DNA conformation in this system is protein-DNA interaction, and not DNA sequence.

Original languageEnglish (US)
Pages (from-to)63-74
Number of pages12
JournalJournal of molecular biology
Volume314
Issue number1
DOIs
StatePublished - Nov 16 2001

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Molecular Biology

Keywords

  • Catabolite activator protein (CAP)
  • Indirect readout
  • Protein-DNA interaction
  • Protein-induced DNA bending
  • cAMP receptor protein (CRP)

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