Mutational analysis of integrase arm-type binding sites of bacteriophage lambda. Integration and excision involve distinct interactions of integrase with arm-type sites

Carl E. Bauer, Steven D. Hesse, Richard I. Gumport, Jeffrey F. Gardner

Research output: Contribution to journalArticlepeer-review

Abstract

Integrative recombination between specific attachment (att) regions of the bacteriophage lambda genome (attP) and the Escherichia coli genome (attB) results in a prophage flanked by the hybrid recombinant sites attL and attR. Each att site contains sequences to which proteins involved in recombination bind. Using site-directed mutagenesis, we have constructed a related set of point mutations within each of the five Int "arm-type" binding sites located within attP, attL and attR. Footprint analyses of binding demonstrate that mutating the arm-type sites significantly disrupts the binding of Int. Recombination analyses of mutant att sites in vivo and in vitro demonstrate that only three wild-type arm type sites within attP are required for efficient integrative recombination. Similar analyses demonstrate that efficient excision can occur with two other different sets of wild-type arm type sites in attL and attR. These results demonstrate that integrative and excisive recombination may involve interactions of Int with distinct and different subsets of arm type sites.

Original languageEnglish (US)
Pages (from-to)513-527
Number of pages15
JournalJournal of Molecular Biology
Volume192
Issue number3
DOIs
StatePublished - Dec 5 1986

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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