The solution structure of Lac repressor headpiece 62 complexed to a symmetrical lac operator

Christian A.E.M. Spronk, Alexandre M.J.J. Bonvin, Plachikkat K. Radha, Giuseppe Melacini, Rolf Boelens, R Kaptein

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Background: Lactose repressor protein (Lac) controls the expression of the lactose metabolic genes in Escherichia coil by binding to an operator sequence in the promoter of the lac operon. Binding of inducer molecules to the Lac core domain induces changes in tertiary structure that are propagated to the DNA-binding domain through the connecting hinge region, thereby reducing the affinity for the operator. Protein-protein and protein-DNA interactions involving the hinge region play a crucial role in the allosteric changes occurring upon induction, but have not, as yet, been analyzed in atomic detail. Results: We have used nuclear magnetic resonance (NMR) spectroscopy and restrained molecular dynamics (rMD) to determine the structure of the Lac repressor DNA-binding domain (headpiece 62; HP62) in complex with a symmetrized lac operator. Analysis of the structures reveals specific interactions between Lac repressor and DNA that were not found in previously investigated Lac repressor-DNA complexes. Important differences with the previously reported structures of the HP56-DNA complex were found in the loop following the helix-turn-helix (HTH) motif. The protein-protein and protein-DNA interactions involving the hinge region and the deformations in the DNA structure could be delineated in atomic detail. The structures were also used for comparison with the available crystallographic data on the Lac and Pur repressor-DNA complexes. Conclusions: The structures of the HP62-DNA complex provide the basis for a better understanding of the specific recognition in the Lac repressor-operator complex. In addition, the structural features of the hinge region provide detailed insight into the protein-protein and protein-DNA interactions responsible for the high affinity of the repressor for operator DNA.
Original languageEnglish
Pages (from-to)1483-1492
Number of pages10
JournalStructure
Volume7
Issue number12
DOIs
Publication statusPublished - 15 Dec 1999

Keywords

  • DNA-induced α helix
  • Lac repressor
  • Molecular dynamics
  • NMR
  • Protein-DNA complex
  • lactose
  • allosterism
  • article
  • crystallography
  • DNA binding
  • DNA protein complex
  • DNA structure
  • Escherichia coli
  • gene expression regulation
  • lactose operon
  • molecular dynamics
  • nuclear magnetic resonance spectroscopy
  • operator gene
  • priority journal
  • promoter region
  • protein DNA interaction
  • protein protein interaction
  • protein structure
  • protein tertiary structure

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