A basic tail increases repression by dimeric lac repressor

J Mol Biol. 1995 Mar 31;247(3):396-403. doi: 10.1006/jmbi.1994.0148.

Abstract

Tetrameric Lac repressor achieves cooperative repression by binding simultaneously to O1 and to one of the auxiliary operators O2 or O3, thereby forcing the intervening DNA into a loop. Dimeric Lac repressor is not able to form DNA loops and consequently shows no cooperative repression. We constructed a dimeric Lac repressor mutant which exhibits increased repression to the lac operon that does not depend on specific operator-repressor-operator loops. This Lac repressor carries a synthetic tail of basic residues attached to its C terminus. With this construct, we observe an increase of the in vivo repression upon addition of auxiliary lac operators to a chromosomal lac operon controlled by O1. This suggests that the basic tail enables dimeric Lac repressor to enhance its repression by additional non-specific DNA contacts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • DNA, Bacterial / metabolism
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Lac Operon / drug effects
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • Operator Regions, Genetic / drug effects*
  • Protein Binding
  • Protein Conformation*
  • Protein Engineering
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Fusion Proteins / pharmacology
  • Repressor Proteins / chemistry*
  • Repressor Proteins / metabolism
  • Repressor Proteins / pharmacology
  • Structure-Activity Relationship

Substances

  • DNA, Bacterial
  • Recombinant Fusion Proteins
  • Repressor Proteins