title: DNA2 cooperates with the WRN and BLM RecQ helicases to mediate long-range DNA end resection in human cells.
abstract: The 5'-3' resection of DNA ends is a prerequisite for the repair of DNA double strand breaks by homologous recombination, microhomology-mediated end joining, and single strand annealing. Recent studies in yeast have shown that, following initial DNA end processing by the Mre11-Rad50-Xrs2 complex and Sae2, the extension of resection tracts is mediated either by exonuclease 1 or by combined activities of the RecQ family DNA helicase Sgs1 and the helicase/endonuclease Dna2. Although human DNA2 has been shown to cooperate with the BLM helicase to catalyze the resection of DNA ends, it remains a matter of debate whether another human RecQ helicase, WRN, can substitute for BLM in DNA2-catalyzed resection. Here we present evidence that WRN and BLM act epistatically with DNA2 to promote the long-range resection of double strand break ends in human cells. Our biochemical experiments show that WRN and DNA2 interact physically and coordinate their enzymatic activities to mediate 5'-3' DNA end resection in a reaction dependent on RPA. In addition, we present in vitro and in vivo data suggesting that BLM promotes DNA end resection as part of the BLM-TOPOIIIα-RMI1-RMI2 complex. Our study provides new mechanistic insights into the process of DNA end resection in mammalian cells.
authors.affiliation: Institute of Molecular Cancer Research, University of Zurich, 8057 Zurich, Switzerland and.
authors.forename: Andreas
authors.initials: A
authors.lastname: Sturzenegger
authors.affiliation: Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, 14300 Prague, Czech Republic.
authors.forename: Kamila
authors.initials: K
authors.lastname: Burdova
authors.affiliation: Institute of Molecular Cancer Research, University of Zurich, 8057 Zurich, Switzerland and.
authors.forename: Radhakrishnan
authors.initials: R
authors.lastname: Kanagaraj
authors.affiliation: Institute of Molecular Cancer Research, University of Zurich, 8057 Zurich, Switzerland and.
authors.forename: Maryna
authors.initials: M
authors.lastname: Levikova
authors.affiliation: Institute of Molecular Cancer Research, University of Zurich, 8057 Zurich, Switzerland and.
authors.forename: Cosimo
authors.initials: C
authors.lastname: Pinto
authors.affiliation: Institute of Molecular Cancer Research, University of Zurich, 8057 Zurich, Switzerland and.
authors.forename: Petr
authors.initials: P
authors.lastname: Cejka
authors.affiliation: Institute of Molecular Cancer Research, University of Zurich, 8057 Zurich, Switzerland and; Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, 14300 Prague, Czech Republic. Electronic address: pjanscak@imcr.uzh.ch.
authors.forename: Pavel
authors.initials: P
authors.lastname: Janscak
mesh_terms: D017766:Acid Anhydride Hydrolases; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D053903:DNA Breaks, Double-Stranded; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D045643:DNA Repair Enzymes / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004843:Epistasis, Genetic / Q000502:physiology*; D005090:Exodeoxyribonucleases / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D000076228:MRE11 Homologue Protein; D009097:Multienzyme Complexes / Q000235:genetics / Q000378:metabolism; D053484:RecQ Helicases / Q000235:genetics / Q000378:metabolism*; D044764:Ubiquitin-Activating Enzymes / Q000235:genetics / Q000378:metabolism; D000071657:Werner Syndrome Helicase
chemical_list: D004268:DNA-Binding Proteins; C121766:MRE11 protein, human; D009097:Multienzyme Complexes; C470301:UBA2 protein, human; D004247:DNA; D005090:Exodeoxyribonucleases; D000076228:MRE11 Homologue Protein; D017766:Acid Anhydride Hydrolases; C112158:RAD50 protein, human; C097151:Bloom syndrome protein; D004265:DNA Helicases; C509908:DNA2 protein, human; D053484:RecQ Helicases; C098785:WRN protein, human; D000071657:Werner Syndrome Helicase; D044764:Ubiquitin-Activating Enzymes; D045643:DNA Repair Enzymes
keywords: DNA Damage; DNA Helicase; DNA Recombination; DNA Repair; Genomic Instability; RecQ