MRE11 is essential for the long-term viability of undifferentiated spermatogonia

Abstract

In the meiotic prophase, programmed SPO11-linked DNA double-strand breaks (DSBs) are repaired by homologous recombination (HR). The MRE11-RAD50-NBS1 (MRN) complex is essential for initiating DNA end resection, the first step of HR. However, residual DNA end resection still occurs in Nbs1 knockout (KO) spermatocytes for unknown reasons. Here, we show that DNA end resection is completely abolished in Mre11 KO spermatocytes. In addition, Mre11 KO, but not Nbs1 KO, undifferentiated spermatogonia are rapidly exhausted due to DSB accumulation, proliferation defects, and elevated apoptosis. Cellular studies reveal that a small amount of MRE11 retained in the nucleus of Nbs1 KO cells likely underlies the differences between Mre11 and Nbs1 KO cells. Taken together, our study not only demonstrates an irreplaceable role of the MRE11 in DNA end resection at SPO11-linked DSBs but also unveils a unique function of MRE11 in maintaining the long-term viability of undifferentiated spermatogonia.

References

[1] ScullyR, PandayA, ElangoR, Willis NA. DNA double-strand break repair-pathway choice in somatic mammalian cells. Nat Rev Mol Cell Biol. 2019; 20(11):698-714.

[2] HerJ, Bunting SF. How cells ensure correct repair of DNA double-strand breaks. J Biol Chem. 2018; 293(27):10502-10511.

[3] SymingtonLS, Gautier J. Double-strand break end resection and repair pathway choice. Annu Rev Genet. 2011; 45:247-271.

[4] HopfnerKP, Karcher A, CraigL, WooTT, CarneyJP, TainerJA. Structural biochemistry and interaction architecture of the DNA double-strand break repair Mre11 nuclease and Rad50-ATPase. Cell. 2001; 105(4):473-485.

[5] CejkaP, Symington LS. DNA end resection: mechanism and control. Annu Rev Genet. 2021; 55:285-307.

[6] HoaNN, Shimizu T, ZhouZW, et al. Mre11 is essential for the removal of lethal topoisomerase 2 covalent cleavage complexes. Mol Cell. 2016; 64(3):580-592.

[7] LamI, KeeneyS. Mechanism and regulation of meiotic recombination initiation. Cold Spring Harb Perspect Biol. 2014; 7(1):a016634.

[8] MimitouEP, Symington LS. DNA end resection: many nucleases make light work. DNA Repair (Amst). 2009; 8(9):983-995.

[9] ZhangB, TangZ, LiL, LuLY. NBS1 is required for SPO11-linked DNA double-strand break repair in male meiosis. Cell Death Differ. 2020; 27(7):2176-2190.

[10] CejkaP. DNA end resection: nucleases team up with the right partners to initiate homologous recombination. J Biol Chem. 2015; 290(38):22931-22938.

[11] CarneyJP, MaserRS, OlivaresH, et al. The hMre11/hRad50 protein complex and Nijmegen breakage syndrome: linkage of double-strand break repair to the cellular DNA damage response. Cell. 1998; 93(3):477-486.

[12] Desai-MehtaA, Cerosaletti KM, ConcannonP. Distinct functional domains of nibrin mediate Mre11 binding, focus formation, and nuclear localization. Mol Cell Biol. 2001; 21(6):2184-2191.

[13] TheunissenJW, KaplanMI, HuntPA, et al. Checkpoint failure and chromosomal instability without lymphomagenesis in Mre11(ATLD1/ATLD1) mice. Mol Cell. 2003; 12(6):1511-1523.

[14] CherrySM, Adelman CA, TheunissenJW, HassoldTJ, HuntPA, PetriniJHJ. The Mre11 complex influences DNA repair, synapsis, and crossing over in murine meiosis. Curr Biol. 2007; 17(4):373-378.

[15] YuZ, VogelG, CoulombeY, et al. The MRE11 GAR motif regulates DNA double-strand break processing and ATR activation. Cell Res. 2012; 22(2):305-320.

[16] ToyookaY, Tsunekawa N, TakahashiY, MatsuiY, SatohM, NoceT. Expression and intracellular localization of mouse vasa-homologue protein during germ cell development. Mech Dev. 2000; 93(1–2):139-149.

[17] HafnerA, BulykML, JambhekarA, Lahav G. The multiple mechanisms that regulate p53 activity and cell fate. Nat Rev Mol Cell Biol. 2019; 20(4):199-210.

[18] DingM, QingX, ZhangG, et al. The essential DNA damage response complex MRN is dispensable for the survival and function of Purkinje neurons. Front Aging Neurosci. 2021; 13:786199.

[19] MimitouEP, Symington LS. Sae2, Exo1 and Sgs1 collaborate in DNA double-strand break processing. Nature. 2008; 455(7214):770-774.

[20] ZhuZ, ChungWH, ShimEY, Lee SE, IraG. Sgs1 helicase and two nucleases Dna2 and Exo1 resect DNA double-strand break ends. Cell. 2008; 134(6):981-994.

[21] PaudyalSC, LiS, YanH, HunterT, YouZ. Dna2 initiates resection at clean DNA double-strand breaks. Nucleic Acids Res. 2017; 45(20):11766-11781.

[22] NimonkarAV, Genschel J, KinoshitaE, et al. BLM-DNA2-RPA-MRN and EXO1-BLM-RPA-MRN constitute two DNA end resection machineries for human DNA break repair. Genes Dev. 2011; 25(4):350-362.

[23] CannavoE, CejkaP, KowalczykowskiSC. Relationship of DNA degradation by Saccharomyces cerevisiae exonuclease 1 and its stimulation by RPA and Mre11-Rad50-Xrs2 to DNA end resection. Proc Natl Acad Sci U S A. 2013; 110(18):E1661-E1668.

[24] NealeMJ, PanJ, KeeneyS. Endonucleolytic processing of covalent protein-linked DNA double-strand breaks. Nature. 2005; 436(7053):1053-1057.

[25] CostanzoV, Robertson K, BibikovaM, et al. Mre11 protein complex prevents double-strand break accumulation during chromosomal DNA replication. Mol Cell. 2001; 8(1):137-147.

[26] BruhnC, ZhouZW, AiH, WangZQ. The essential function of the MRN complex in the resolution of endogenous replication intermediates. Cell Rep. 2014; 6(1):182-195.

[27] StewartGS, MaserRS, StankovicT, et al. The DNA double-strand break repair gene hMRE11 is mutated in individuals with an ataxia-telangiectasia-like disorder. Cell. 1999; 99(6):577-587.

[28] VaronR, Vissinga C, PlatzerM, et al. Nibrin, a novel DNA double-strand break repair protein, is mutated in Nijmegen breakage syndrome. Cell. 1998; 93(3):467-476.

[29] TaylorAMR, Rothblum-Oviatt C, EllisNA, et al. Chromosome instability syndromes. Nat Rev Dis Primers. 2019; 5(1):64.

[30] WilliamsBR, Mirzoeva OK, MorganWF, LinJ, Dunnick W, PetriniJHJ. A murine model of Nijmegen breakage syndrome. Curr Biol. 2002; 12(8):648-653.

[31] BuisJ, WuY, DengY, et al. Mre11 nuclease activity has essential roles in DNA repair and genomic stability distinct from ATM activation. Cell. 2008; 135(1):85-96.

[32] LuoG, YaoMS, BenderCF, et al. Disruption of mRad50 causes embryonic stem cell lethality, abnormal embryonic development, and sensitivity to ionizing radiation. Proc Natl Acad Sci U S A. 1999; 96(13):7376-7381.

[33] ZhuJ, Petersen S, TessarolloL, NussenzweigA. Targeted disruption of the Nijmegen breakage syndrome gene NBS1 leads to early embryonic lethality in mice. Curr Biol. 2001; 11(2):105-109.

[34] XiaoY, WeaverDT. Conditional gene targeted deletion by Cre recombinase demonstrates the requirement for the double-strand break repair Mre11 protein in murine embryonic stem cells. Nucleic Acids Res. 1997; 25(15):2985-2991.

[35] YangYG, SaidiA, FrappartPO, et al. Conditional deletion of Nbs1 in murine cells reveals its role in branching repair pathways of DNA double-strand breaks. EMBO J. 2006; 25(23):5527-5538.

RIGHTS & PERMISSIONS

2024 The Author(s). Cell Proliferation published by Beijing Institute for Stem Cell and Regenerative Medicine and John Wiley & Sons Ltd.