Role of Escherichia coli IHF protein in lambda site-specific recombination. A mutational analysis of binding sites

Jeffrey F. Gardner, Howard A. Nash

Research output: Contribution to journalArticle

Abstract

The phage lambda attachment site, attP, contains three binding sites for an Escherichia coli protein, IHF, that is needed for efficient integrative recombination. We have used synthetic oligodeoxyribonuleotides to direct multiple base changes at each of these three sites. Alteration by two base-pairs of the consensus sequence for the leftmost binding site specifically interferes with IHF binding to that site and modestly depresses recombination in vitro. For each of the three binding sites, alteration of the consensus sequence by four base-pairs strongly depresses recombination in vitro, indicating that all three sites are important for attP function. The mutated attP sites are also depressed for recombination in vivo but some of the mutants are less affected than they are in vitro. The disparity between effects in vivo and in vitro for some mutants but not others suggests that the three binding sites are not functionally equivalent and that at some sites additional E. coli factors may replace or assist IHF. The non-equivalence of the three IHF sites is also indicated by the behavior of prophage attachment sites carrying mutations in the binding sites.

Original languageEnglish (US)
Pages (from-to)181-189
Number of pages9
JournalJournal of Molecular Biology
Volume191
Issue number2
DOIs
StatePublished - Sep 20 1986
Externally publishedYes

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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