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DNA2 cooperates with the WRN and BLM RecQ helicases to mediate long-range DNA-end resection in human cells


Sturzenegger, Andreas; Burdova, Kamila; Kanagaraj, Radhakrishnan; Levikova, Maryna; Pinto, Cosimo; Cejka, Petr; Janscak, Pavel (2014). DNA2 cooperates with the WRN and BLM RecQ helicases to mediate long-range DNA-end resection in human cells. Journal of Biological Chemistry, 289(39):27314-27326.

Abstract

The 5'-3' resection of DNA ends is a prerequisite for 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 an initial DNA-end processing by the Mre11-Rad50-Xrs2 complex and Sae2, 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 was shown to cooperate with the BLM helicase to catalyze resection of DNA ends, it remains a matter of debate whether another human RecQ helicase, namely WRN, can substitute for BLM in DNA2-catalyzed resection. Here, we present evidence that WRN and BLM act epistatically with DNA2 to promote long-range resection of DSB 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.

Abstract

The 5'-3' resection of DNA ends is a prerequisite for 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 an initial DNA-end processing by the Mre11-Rad50-Xrs2 complex and Sae2, 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 was shown to cooperate with the BLM helicase to catalyze resection of DNA ends, it remains a matter of debate whether another human RecQ helicase, namely WRN, can substitute for BLM in DNA2-catalyzed resection. Here, we present evidence that WRN and BLM act epistatically with DNA2 to promote long-range resection of DSB 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.

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Item Type:Journal Article, refereed, original work
Communities & Collections:04 Faculty of Medicine > Institute of Molecular Cancer Research
07 Faculty of Science > Institute of Molecular Cancer Research
Dewey Decimal Classification:570 Life sciences; biology
Language:English
Date:13 August 2014
Deposited On:30 Sep 2014 15:17
Last Modified:08 Dec 2017 07:12
Publisher:American Society for Biochemistry and Molecular Biology
ISSN:0021-9258
Additional Information:This research was originally published in Journal Name.Sturzenegger A et al: DNA2 Cooperates with the WRN and BLM RecQ Helicases to Mediate Long-range DNA End Resection in Human Cells. The Journal of Biological Chemistry, 2014, 289:27314-27326 © the American Society for Biochemistry and Molecular Biology.
Free access at:PubMed ID. An embargo period may apply.
Publisher DOI:https://doi.org/10.1074/jbc.M114.578823
PubMed ID:25122754

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