title: VCP/p97 Extracts Sterically Trapped Ku70/80 Rings from DNA in Double-Strand Break Repair.
abstract: During DNA double-strand break (DSB) repair, the ring-shaped Ku70/80 complex becomes trapped on DNA and needs to be actively extracted, but it has remained unclear what provides the required energy. By means of reconstitution of DSB repair on beads, we demonstrate here that DNA-locked Ku rings are released by the AAA-ATPase p97. To achieve this, p97 requires ATP hydrolysis, cooperates with the Ufd1-Npl4 ubiquitin-adaptor complex, and specifically targets Ku80 that is modified by K48-linked ubiquitin chains. In U2OS cells, chemical inhibition of p97 or siRNA-mediated depletion of p97 or its adapters impairs Ku80 removal after non-homologous end joining of DSBs. Moreover, this inhibition attenuates early steps in homologous recombination, consistent with p97-driven Ku release also affecting repair pathway choice. Thus, our data answer a central question regarding regulation of Ku in DSB repair and illustrate the ability of p97 to segregate even tightly bound protein complexes for release from DNA.
authors.affiliation: Molecular Biology I, Faculty of Biology, University of Duisburg-Essen, 45117 Essen, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: van den Boom
authors.affiliation: Molecular Biology I, Faculty of Biology, University of Duisburg-Essen, 45117 Essen, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Wolf
authors.affiliation: Molecular Biology I, Faculty of Biology, University of Duisburg-Essen, 45117 Essen, Germany.
authors.forename: Lena
authors.initials: L
authors.lastname: Weimann
authors.affiliation: ICCE, Faculty of Biology, University of Duisburg-Essen, 45117 Essen, Germany.
authors.forename: Nina
authors.initials: N
authors.lastname: Schulze
authors.affiliation: Institute of Medical Radiation Biology, Medical School, University of Duisburg-Essen, 45122 Essen, Germany.
authors.forename: Fanghua
authors.initials: F
authors.lastname: Li
authors.affiliation: Chemical Proteomics, Faculty of Biology, University of Duisburg-Essen, 45117 Essen, Germany.
authors.forename: Farnusch
authors.initials: F
authors.lastname: Kaschani
authors.affiliation: Molecular Biology I, Faculty of Biology, University of Duisburg-Essen, 45117 Essen, Germany.
authors.forename: Anne
authors.initials: A
authors.lastname: Riemer
authors.affiliation: Laboratory of Chromosome and Cell Biology, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Christian
authors.initials: C
authors.lastname: Zierhut
authors.affiliation: Chemical Proteomics, Faculty of Biology, University of Duisburg-Essen, 45117 Essen, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Kaiser
authors.affiliation: Institute of Medical Radiation Biology, Medical School, University of Duisburg-Essen, 45122 Essen, Germany.
authors.forename: George
authors.initials: G
authors.lastname: Iliakis
authors.affiliation: Laboratory of Chromosome and Cell Biology, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Hironori
authors.initials: H
authors.lastname: Funabiki
authors.affiliation: Molecular Biology I, Faculty of Biology, University of Duisburg-Essen, 45117 Essen, Germany. Electronic address: hemmo.meyer@uni-due.de.
authors.forename: Hemmo
authors.initials: H
authors.lastname: Meyer
mesh_terms: D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D029845:Amphibian Proteins / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D004247:DNA / Q000235:genetics / Q000378:metabolism; D053903:DNA Breaks, Double-Stranded; D059766:DNA End-Joining Repair; D005786:Gene Expression Regulation; D006801:Humans; D006868:Hydrolysis; D000072200:Ku Autoantigen / Q000235:genetics* / Q000378:metabolism; D008863:Microspheres; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism*; D010063:Ovum / Q000737:chemistry / Q000166:cytology; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D059767:Recombinational DNA Repair; D000074405:Valosin Containing Protein; D014982:Xenopus laevis
chemical_list: D029845:Amphibian Proteins; D018797:Cell Cycle Proteins; D034741:RNA, Small Interfering; D000255:Adenosine Triphosphate; D004247:DNA; D000251:Adenosine Triphosphatases; C000614178:VCP protein, human; D000074405:Valosin Containing Protein; D000072200:Ku Autoantigen
keywords: DNA repair; Ku; Ku80; NHEJ; VCP; chromatin; double strand break; homologous recombination; p97