title: Genetic analysis of the bacteriophage lambda attL nucleoprotein complex.
abstract: Site-specific recombination in bacteriophage lambda involves interactions among proteins required for integration and excision of DNA molecules. We have analyzed the elements required to form an in vivo nucleoprotein complex of integrase (Int) and integration host factor (IHF). Interaction of Int with the core (the site of strand exchange) is stabilized by the flanking arm region of attL. IHF, in addition to Int, is required for efficient Int-core binding. We used the in vivo attL binding assay to characterize several Int variants for their abilities to form stable attL complexes. Substitution of Int active site tyrosine 342 by phenylalanine had no effect on the ability of the protein to form attL complexes. Three other amino acids that are completely conserved in the integrase family of recombinases (arginine 212, histidine 308, and arginine 311) were separately substituted by glutamine, leucine, and histidine, respectively. In each case, the mutant protein was altered in its ability to form attL complexes while retaining its ability to bind to the lambda arm-type sites. We propose that, in addition to their role in catalysis, this triad of amino acids helps the Int protein to interact with the lambda core sites.
authors.affiliation: Department of Microbiology, University of Illinois, Urbana 61801, USA.
authors.forename: M P
authors.initials: MP
authors.lastname: MacWilliams
authors.forename: R I
authors.initials: RI
authors.lastname: Gumport
authors.forename: J F
authors.initials: JF
authors.lastname: Gardner
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D010582:Bacteriophage lambda / Q000235:genetics*; D001665:Binding Sites; D034982:Integration Host Factors; D016296:Mutagenesis; D009698:Nucleoproteins / Q000235:genetics*; D012097:Repressor Proteins / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics*; D054334:Viral Regulatory and Accessory Proteins
chemical_list: D001426:Bacterial Proteins; D034982:Integration Host Factors; C059417:N protein, Bacteriophage lambda; D009698:Nucleoproteins; D012097:Repressor Proteins; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins