Stepwise 5' DNA end-specific resection of DNA breaks by the Mre11-Rad50-Xrs2 and Sae2 nuclease ensemble

Proceedings of the National Academy of Sciences of the United States of America
Elda CannavoPetr Cejka

Abstract

To repair DNA double-strand breaks by homologous recombination, the 5'-terminated DNA strands must first be resected to produce 3' overhangs. Mre11 from Saccharomyces cerevisiae is a 3' → 5' exonuclease that is responsible for 5' end degradation in vivo. Using plasmid-length DNA substrates and purified recombinant proteins, we show that the combined exonuclease and endonuclease activities of recombinant MRX-Sae2 preferentially degrade the 5'-terminated DNA strand, which extends beyond the vicinity of the DNA end. Mechanistically, Rad50 restricts the Mre11 exonuclease in an ATP binding-dependent manner, preventing 3' end degradation. Phosphorylated Sae2, along with stimulating the MRX endonuclease as shown previously, also overcomes this inhibition to promote the 3' → 5' exonuclease of MRX, which requires ATP hydrolysis by Rad50. Our results support a model in which MRX-Sae2 catalyzes 5'-DNA end degradation by stepwise endonucleolytic DNA incisions, followed by exonucleolytic 3' → 5' degradation of the individual DNA fragments. This model explains how both exonuclease and endonuclease activities of Mre11 functionally integrate within the MRX-Sae2 ensemble to resect 5'-terminated DNA.

References

Nov 21, 1995·Proceedings of the National Academy of Sciences of the United States of America·S Keeney, N Kleckner
Nov 19, 1993·Cell·L L Sandell, V A Zakian
Aug 15, 1998·The Journal of Biological Chemistry·K M TrujilloP Sung
May 24, 2000·Proceedings of the National Academy of Sciences of the United States of America·T T Paull, M Gellert
Jul 17, 2001·The Journal of Biological Chemistry·K M Trujillo, P Sung
Dec 14, 2001·Molecular Cell·M de JagerC Wyman
Oct 16, 2004·Molecular and Cellular Biology·Bertrand Llorente, Lorraine S Symington
Nov 18, 2004·The Journal of Biological Chemistry·Ling ChenAlan E Tomkinson
Sep 16, 2005·The Journal of Biological Chemistry·Michela ClericiMaria Pia Longhese
Aug 22, 2008·Nature·Pablo HuertasStephen P Jackson
Sep 23, 2008·Nature·Eleni P Mimitou, Lorraine S Symington
Oct 23, 2010·Molecular Cell·Alberto Ciccia, Stephen J Elledge
Aug 10, 2011·Proceedings of the National Academy of Sciences of the United States of America·Michael L Oldham, Jue Chen
Aug 31, 2011·Molecular and Cellular Biology·Steven S FosterJohn H J Petrini
Oct 18, 2011·Nature·Valerie GarciaMatthew J Neale
Nov 22, 2011·The Journal of Biological Chemistry·Jerzy MajkaCynthia T McMurray
Apr 17, 2013·Proceedings of the National Academy of Sciences of the United States of America·Elda CannavoStephen C Kowalczykowski
Jun 4, 2013·Nature Structural & Molecular Biology·Nicholas L AdkinsCraig L Peterson
Oct 18, 2014·Cold Spring Harbor Perspectives in Biology·Isabel Lam, Scott Keeney
Nov 4, 2015·Cold Spring Harbor Perspectives in Biology·Stephen C Kowalczykowski
Oct 22, 2016·Molecular Cell·Julyun OhLorraine S Symington
Jan 7, 2017·Science·Eleni P MimitouScott Keeney
May 18, 2017·Nature Reviews. Molecular Cell Biology·Howard H Y ChangMichael R Lieber

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Citations

Aug 12, 2020·Journal of Radiation Research·Atsushi Shibata, Penny A Jeggo
Jun 25, 2019·Frontiers in Molecular Biosciences·Erika CasariMaria Pia Longhese
Mar 29, 2020·Biochemical Society Transactions·Chiara Vittoria ColomboMaria Pia Longhese
Oct 14, 2020·Proceedings of the National Academy of Sciences of the United States of America·Nicole StantialJohn L Nitiss
Jun 4, 2020·Computational and Structural Biotechnology Journal·Renata TisiMaria Pia Longhese
Jan 16, 2021·Journal of Zhejiang University. Science. B·Shan Qiu, Jun Huang
Jun 2, 2020·DNA Repair·Giordano Reginato, Petr Cejka
Apr 11, 2021·Proceedings of the National Academy of Sciences of the United States of America·Aleksandar ZdravkovićHiroshi Iwasaki
May 3, 2021·The Journal of Biological Chemistry·Andrew R YoblinskiRaghuvar Dronamraju
May 14, 2021·Nature Communications·Florian Roisné-HamelinStéphane Marcand
Jun 24, 2021·Nature Communications·Freddy KhayatAlessandro Bianchi
Jul 30, 2021·Frontiers in Cell and Developmental Biology·Stephanie M AckersonJason A Stewart
Jul 31, 2021·Current Opinion in Genetics & Development·Robert Gnügge, Lorraine S Symington
Jul 27, 2021·Current Opinion in Genetics & Development·Diego BonettiMaria Pia Longhese
Nov 24, 2021·Annual Review of Genetics·Petr Cejka, Lorraine S Symington

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