Control of prophage integration and excision in bacteriophage P2: nucleotide sequences of the int gene and att sites

Abstract

Integration of bacteriophage P2 into the Escherichia coli host genome involves recombination between two specific attachment sites, attP and attB, one on the phage and the other on the host genome, respectively. The reaction is controlled by the product of the phage int gene, a basic polypeptide of about 37 kDa [Ljungquist and Bertani, Mol.Gen.Genet.192(1983) 87–94].The int gene appears to be expressed differently by an infecting phage, as opposed to a prophage[Bertani, Proc.Natl.Acad.Sci.USA 65 (1970) 331–336].A 1200-bp region of P2 DNA containing the int gene and attP,the prophage hybrid ends attL and attR,and one bacterial attachment site,the preferred site locIfrom E.coli strain C, have all been sequenced. An open reading frame coding for a polypeptide of 337 amino acids corresponds to the int gene. The gene has no obvious promoter sequence preceding it. The int gene transcript seems to continue past the attP site downstream from it, suggesting a possible explanation for the previously observed difference in integration and excision.A comparison of the four attachment sites reveals a common ‘core’ sequence of 27 bp: 5'-AAAAAATAAGCCCGTGTAAGGGAGATT-3'.The P2 nip1 mutation, which increases prophage excision [Calendar et al.,Virology 47 (1972) 68–75], was found to lie within the int gene itself. The P2 saf variant,which has altered site preference [Six, Virology 29 (1966) 106–125], has a bp substitution within the core sequence.Three deletion/substitution mutants, vir22, vir94 and del3, also have altered core sequences.