COMPLETE NUCLEOTIDE-SEQUENCE AND MOLECULAR CHARACTERIZATION OF THE TEMPERATE STAPHYLOCOCCAL BACTERIOPHAGE PHI-PVL CARRYING PANTON-VALENTINELEUKOCIDIN GENES

The staphylococcal Panton-Valentine leukocidin (PVL) genes, [lukS-PV-l ukF-PV], exist on the genome of a temperate bacteriophage phi PVL isol ated from mitomycin C-induced Staphylococcus aureus V8 (ATCC 49775) (K aneko, J., Kimura, T., Kawakami, Y., Tomita, T., Kamio, Y., 1997b. Pan ton-Valentine leukocidin genes in phage-like particle isolated from mi tomycin C-treated Staphylococcus aureus V8 (ATCC 49775). Biosci. Biote chnol. Biochem. 61, 1960-1962). In this study, the complete nucleotide sequence of the phi PVL genome was analyzed, and the att sites (attL, attR, and attB) required for site-specific integration of phi PVL int o the host chromosome were also determined. The linear double-stranded phi PVL genome comprised 41 401 bp with 3' staggered cohesive ends (c os) of nine bases and contained 63 ORFs, among which the regulatory pr oteins involved in DNA replication, structural proteins, a holin, a ly sin, an integrase, and dUTPase, were tentatively identified by the com parison of the deduced amino acid sequences and by the analysis of the proteins isolated from phi PVL particles. The [lukS-PV-lukF-PV], attP , and int (integrase gene) of phi PVL were all located very close to o ne another within a 4.0-kb segment on the genome in the order given, a nd this segment was located at the center from the left and the right cos sites. In addition, the attP region contained five direct repeat s equences that showed a high degree of homology with the recombinase-bi nding sites of some other S. aureus bacteriophages. The phi PVL genome was found to integrate into an ORF encoding an unknown protein compri sing 725 amino acid residues with two leucine zipper-like moths. (C) 1 998 Elsevier Science B.V. All rights reserved.