title: Attachment site selection and identity in Bxb1 serine integrase-mediated site-specific recombination.
abstract: Phage-encoded serine integrases mediate directionally regulated site-specific recombination between short attP and attB DNA sites without host factor requirements. These features make them attractive for genome engineering and synthetic genetics, although the basis for DNA site selection is poorly understood. Here we show that attP selection is determined through multiple proofreading steps that reject non-attP substrates, and that discrimination of attP and attB involves two critical site features: the outermost 5-6 base pairs of attP that are required for Int binding and recombination but antagonize attB function, and the "discriminators" at positions -15/+15 that determine attB identity but also antagonize attP function. Thus, although the attachment sites differ in length and sequence, only two base changes are needed to convert attP to attL, and just two more from attL to attB. The opposing effect of site identifiers ensures that site schizophrenia with dual identities does not occur.
authors: Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors: Shweta
authors: S
authors: Singh
authors: Pallavi
authors: P
authors: Ghosh
authors: Graham F
authors: GF
authors: Hatfull
mesh_terms: D001287:Attachment Sites, Microbiological / Q000235:genetics*; D020029:Base Pairing; D004247:DNA / Q000235:genetics; D004254:DNA Nucleotidyltransferases / Q000235:genetics* / Q000378:metabolism; D009160:Mycobacteriophages / Q000235:genetics*; D011995:Recombination, Genetic; D012694:Serine / Q000235:genetics; D016662:Virus Integration / Q000235:genetics
chemical_list: D012694:Serine; D004247:DNA; D004254:DNA Nucleotidyltransferases; C077570:Site-specific recombinase
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