Single-molecule studies of repressor-DNA interactions show long-range interactions

Proc Natl Acad Sci U S A. 2005 Jul 12;102(28):9796-801. doi: 10.1073/pnas.0502917102. Epub 2005 Jun 30.

Abstract

We have performed single-molecule studies of GFP-LacI repressor proteins bound to bacteriophage lambda DNA containing a 256 tandem lac operator insertion confined in nanochannels. An integrated photon molecular counting method was developed to determine the number of proteins bound to DNA. By using this method, we determined the saturated mean occupancy of the 256 tandem lac operators to be 13, which constitutes only 2.5% of the available sites. This low occupancy level suggests that the repressors influence each other even when they are widely separated, at distances on the order of 200 nm, or several DNA persistence lengths.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacteriophage lambda / genetics*
  • Biophysical Phenomena
  • Biophysics
  • Carbocyanines
  • DNA, Viral / metabolism*
  • DNA-Binding Proteins / isolation & purification*
  • Electrophoresis / methods
  • Green Fluorescent Proteins / metabolism
  • Nanotechnology / methods
  • Operator Regions, Genetic / genetics*
  • Repressor Proteins / metabolism*

Substances

  • BOBO 3
  • Carbocyanines
  • DNA, Viral
  • DNA-Binding Proteins
  • Repressor Proteins
  • Green Fluorescent Proteins