Hinge-helix formation and DNA bending in various lac repressor-operator complexes

EMBO J. 1999 Nov 15;18(22):6472-80. doi: 10.1093/emboj/18.22.6472.

Abstract

The hinge-region of the lac repressor plays an important role in the models for induction and DNA looping in the lac operon. When lac repressor is bound to a tight-binding symmetric operator, this region forms an alpha-helix that induces bending of the operator. The presence of the hinge-helices is questioned by previous data that suggest that the repressor does not bend the wild-type operator. We show that in the wild-type complex the hinge-helices are formed and the DNA is bent, similar to the symmetric complex. Furthermore, our data show differences in the binding of the DNA binding domains to the half-sites of the wild-type operator and reveal the role of the central base-pair of the wild-type operator in the repressor-operator interaction. The differences in binding to the operator half-sites are incorporated into a model that explains the relative affinities of the repressor for various lac operator sequences that contain left and right half-sites with different spacer lengths.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / metabolism*
  • Base Sequence
  • Cloning, Molecular
  • Escherichia coli Proteins*
  • Lac Operon*
  • Lac Repressors
  • Nuclear Magnetic Resonance, Biomolecular
  • Nucleic Acid Conformation*
  • Oligodeoxyribonucleotides / chemistry*
  • Protein Conformation
  • Repressor Proteins / chemistry*
  • Repressor Proteins / metabolism*

Substances

  • Bacterial Proteins
  • Escherichia coli Proteins
  • Lac Repressors
  • Oligodeoxyribonucleotides
  • Repressor Proteins