title: Chimeric recombinases with designed DNA sequence recognition.
abstract: Site-specific recombination typically occurs only between DNA sequences that have co-evolved with a natural recombinase enzyme to optimize sequence recognition, catalytic efficiency, and regulation. Here, we show that the sequence recognition and the catalysis functions of a recombinase can be specified by unrelated protein domains. We describe chimeric recombinases with a catalytic domain from an activated multiple mutant of the bacterial enzyme Tn3 resolvase, fused to a DNA recognition domain from the mouse transcription factor Zif268. These proteins catalyze efficient recombination specifically at synthetic target sites recognized by two Zif268 domains. Our results demonstrate the functional autonomy of the resolvase catalytic domain and open the way to creating "custom-built" recombinases that act at chosen natural target sequences.
authors.affiliation: Institute of Biomedical and Life Sciences, University of Glasgow, 56 Dumbarton Road, Glasgow G11 6NU, Scotland.
authors.forename: Aram
authors.initials: A
authors.lastname: Akopian
authors.forename: Jiuya
authors.initials: J
authors.lastname: He
authors.forename: Martin R
authors.initials: MR
authors.lastname: Boocock
authors.forename: W Marshall
authors.initials: WM
authors.lastname: Stark
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D020134:Catalytic Domain / Q000235:genetics; D004254:DNA Nucleotidyltransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D010957:Plasmids / Q000235:genetics; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D045522:Recombinases; D011995:Recombination, Genetic; D019895:Transposases / Q000737:chemistry / Q000378:metabolism; D045564:Transposon Resolvases
chemical_list: D004269:DNA, Bacterial; D011993:Recombinant Fusion Proteins; D045522:Recombinases; D004254:DNA Nucleotidyltransferases; C473491:Tn3 resolvase; D019895:Transposases; D045564:Transposon Resolvases