Structural alterations and conformational dynamics in Holliday junctions induced by binding of a site-specific recombinase.
Binding of a cleavage-incompetent mutant of the Flp recombinase induces a roughly square-planar geometry in synthetic immobile Holliday junctions. The branch points, which are rigidly fixed in these junctions in their free forms, tend to be more flexible in their protein-bound forms. Our results (1) suggest a plausible mechanism for the switching of the recombination complex from the Holliday-forming mode to the Holliday-resolving mode, (2) provide a rationale for previous observations that Flp resolves preformed immobile Holliday structures in the parental or in the recombinant mode in a relatively unbiased manner, and (3) accommodate two modes of DNA cleavage by Flp (transhorizontal or transdiagonal) in Holliday substrates.
Lee
J
J
Department of Microbiology, University of Texas at Austin 78712, USA.
Voziyanov
Y
Y
Pathania
S
S
Jayaram
M
M
D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D010582:Bacteriophage lambda / Q000378:metabolism; D004247:DNA / Q000737:chemistry; D004254:DNA Nucleotidyltransferases / Q000737:chemistry* / Q000378:metabolism*; D004586:Electrophoresis; D004706:Endodeoxyribonucleases / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D045565:Holliday Junction Resolvases; D019426:Integrases / Q000378:metabolism; D009690:Nucleic Acid Conformation; D011485:Protein Binding / Q000235:genetics; D011487:Protein Conformation; D045522:Recombinases; D011995:Recombination, Genetic / Q000502:physiology*; D013379:Substrate Specificity; D014764:Viral Proteins / Q000378:metabolism
D001425:Bacterial Outer Membrane Proteins; D045522:Recombinases; D014764:Viral Proteins; D004247:DNA; D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase; D019426:Integrases; C115006:integron integrase IntI1; D004706:Endodeoxyribonucleases; D045565:Holliday Junction Resolvases