title: The orientation of mycobacteriophage Bxb1 integration is solely dependent on the central dinucleotide of attP and attB.
abstract: Integration of the mycobacteriophage Bxb1 genome into its host chromosome is catalyzed by a serine-integrase, a member of the transposon-resolvase family of site-specific recombinases. These enzymes use a concerted mechanism of strand exchange involving double-stranded cleavages with two-base extensions, and covalent protein-DNA linkages via phosphoserine bonds. In contrast to the resolvase/invertase recombination systems--where there are strict requirements for a specific synaptic complex within which the catalytic potential of the enzyme is activated--synapsis of attP and attB by Bxb1 integrase is completely promiscuous, aligning the sites with equal proclivity in parallel and antiparallel alignments. Moreover, the catalytic potential of Bxb1 integrase is fully active in either alignment. As a consequence, the nonpalindromic central dinucleotide (5'-GT) at the center of attP and attB is the sole determinant of Bxb1 prophage orientation, and a single base pair substitution in the two sites is sufficient to eliminate orientation control.
authors.affiliation: Pittsburgh Bacteriophage Institute and Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260, USA.
authors.forename: Pallavi
authors.initials: P
authors.lastname: Ghosh
authors.forename: Amy I
authors.initials: AI
authors.lastname: Kim
authors.forename: Graham F
authors.initials: GF
authors.lastname: Hatfull
mesh_terms: D001287:Attachment Sites, Microbiological; D020029:Base Pairing; D001483:Base Sequence; D026942:DNA Topoisomerases / Q000378:metabolism; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism*; D019426:Integrases / Q000378:metabolism; D008242:Lysogeny; D009160:Mycobacteriophages / Q000235:genetics* / Q000378:metabolism; D011995:Recombination, Genetic; D014764:Viral Proteins / Q000378:metabolism
chemical_list: D004279:DNA, Viral; D014764:Viral Proteins; D019426:Integrases; D026942:DNA Topoisomerases