DNA sequence determinant for FIp-induced DNA bending.
The Flp site-specific recombinase from Saccharomyces cerevisiae induces DNA bending upon interaction with the Flp recognition target (FRT) site. The minimal FRT site comprises the inverted a and b binding elements, which flank a central 8 bp core region. The DNA bend in a complex of two Flp monomers bound to the FRT site is located in the middle of the core region. When the central AT basepair was replaced with a CG, the DNA bend was positioned at the outside end of the core region adjacent to the a binding element. The other basepairs surrounding the central AT basepair were not important to the position of Flp-induced bends. The change also decreased Flp-mediated cleavage of the top strand of the FRT site and increased Flp-mediated cleavage of the bottom strand. The overall recombination proficiency of the site was impaired. We conclude that the central AT basepair provides a point of flexure in the FRT site, which Flp uses to position the bend in dimeric Flp-DNA complexes, and that the structure of the core DNA influences the functionality of the site.
Luetke
K H
KH
Department of Medical Genetics and Microbiology, University of Toronto, Ontario, Canada.
Sadowski
P D
PD
D020029:Base Pairing; D001665:Binding Sites; D004254:DNA Nucleotidyltransferases / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism*; D016296:Mutagenesis; D009154:Mutation; D009838:Oligodeoxyribonucleotides / Q000378:metabolism; D011995:Recombination, Genetic; D017422:Sequence Analysis, DNA
D005656:Fungal Proteins; D009838:Oligodeoxyribonucleotides; D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase