title: ATP-dependent and independent functions of Rad54 in genome maintenance.
abstract: Rad54, a member of the SWI/SNF protein family of DNA-dependent ATPases, repairs DNA double-strand breaks (DSBs) through homologous recombination. Here we demonstrate that Rad54 is required for the timely accumulation of the homologous recombination proteins Rad51 and Brca2 at DSBs. Because replication protein A and Nbs1 accumulation is not affected by Rad54 depletion, Rad54 is downstream of DSB resection. Rad54-mediated Rad51 accumulation does not require Rad54's ATPase activity. Thus, our experiments demonstrate that SWI/SNF proteins may have functions independent of their ATPase activity. However, quantitative real-time analysis of Rad54 focus formation indicates that Rad54's ATPase activity is required for the disassociation of Rad54 from DNA and Rad54 turnover at DSBs. Although the non-DNA-bound fraction of Rad54 reversibly interacts with a focus, independent of its ATPase status, the DNA-bound fraction is immobilized in the absence of ATP hydrolysis by Rad54. Finally, we show that ATP hydrolysis by Rad54 is required for the redistribution of DSB repair sites within the nucleus.
authors.affiliation: Department of Cell Biology and Genetics, Cancer Genomics Center, 3000 CA Rotterdam, Netherlands.
authors.forename: Sheba
authors.initials: S
authors.lastname: Agarwal
authors.forename: Wiggert A
authors.initials: WA
authors.lastname: van Cappellen
authors.forename: Aude
authors.initials: A
authors.lastname: Guénolé
authors.forename: Berina
authors.initials: B
authors.lastname: Eppink
authors.forename: Sam E V
authors.initials: SE
authors.lastname: Linsen
authors.forename: Erik
authors.initials: E
authors.lastname: Meijering
authors.forename: Adriaan
authors.initials: A
authors.lastname: Houtsmuller
authors.forename: Roland
authors.initials: R
authors.lastname: Kanaar
authors.forename: Jeroen
authors.initials: J
authors.lastname: Essers
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism / Q000502:physiology*; D000818:Animals; D002478:Cells, Cultured; D053903:DNA Breaks, Double-Stranded; D004265:DNA Helicases / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D004260:DNA Repair; D016678:Genome; D049452:Green Fluorescent Proteins / Q000032:analysis; D042581:Intranuclear Space / Q000378:metabolism / Q000648:ultrastructure; D051379:Mice; D009687:Nuclear Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D051135:Rad51 Recombinase / Q000032:analysis / Q000378:metabolism / Q000502:physiology; D011995:Recombination, Genetic
chemical_list: D009687:Nuclear Proteins; D049452:Green Fluorescent Proteins; D000255:Adenosine Triphosphate; D051135:Rad51 Recombinase; C495154:Rad51 protein, mouse; D004265:DNA Helicases; C103534:Rad54l protein, mouse