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Related Experiment Videos

A consensus sequence for binding of Lrp to DNA

Y Cui1, Q Wang, G D Stormo

  • 1Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, New York 14853, USA.

Journal of Bacteriology
|September 1, 1995
PubMed
Summary
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Modulation of Lrp action in Escherichia coli by leucine: effects on non-specific binding of Lrp to DNA.

Journal of molecular biology·2001

Leucine-responsive regulatory protein (Lrp) in Escherichia coli binds specific DNA sequences to regulate gene expression. This study identified Lrp binding sites and found that leucine does not alter Lrp

Area of Science:

  • Molecular Biology
  • Genetics
  • Microbiology

Background:

  • Leucine-responsive regulatory protein (Lrp) is a key regulator of gene expression in Escherichia coli.
  • Lrp positively regulates the ilvIH operon by binding to multiple upstream DNA sites.
  • Previous studies proposed a tentative consensus sequence for Lrp binding sites.

Purpose of the Study:

  • To precisely characterize the DNA binding specificity of Lrp.
  • To identify novel Lrp binding sequences using an in vitro selection method.
  • To investigate whether leucine influences Lrp binding specificity.

Main Methods:

  • A variation of the SELEX (systematic evolution of ligands by exponential enrichment) procedure was employed.
  • A pool of 10^12 unique DNA molecules was used to select Lrp-binding sequences.

Related Experiment Videos

  • Binding affinities of selected sequences were estimated, and a new consensus sequence was derived.

    • Main Results:

    • Sixty-three related DNA sequences that bind Lrp were identified.
    • A refined consensus binding sequence (YAGHAWATTWT DCTR) with dyad symmetry was determined.
    • DNA sequences selected with or without leucine were similar, indicating no change in binding specificity.

    Conclusions:

    • The refined consensus sequence provides a more accurate model for Lrp DNA recognition.
    • Leucine does not appear to alter the DNA binding specificity of Lrp.
    • Lrp binding specificity is conserved and not significantly modulated by leucine presence.