In response to genotoxic stress, cells protect their genome integrity by activation of a conserved DNA damage response (DDR) pathway that coordinates DNA repair and progression through the cell cycle. Extensive modification of the chromatin flanking the DNA lesion by ATM kinase and RNF8/RNF168 ubiquitin ligases enables recruitment of various repair factors. Among them BRCA1 and 53BP1 are required for homologous recombination and non-homologous end joining, respectively. Whereas mechanisms of DDR are relatively well understood in interphase cells, comparatively less is known about organization of DDR during mitosis. Although ATM can be activated in mitotic cells, 53BP1 is not recruited to the chromatin until cells exit mitosis. Here we report mitotic phosphorylation of 53BP1 by Plk1 and Cdk1 that impairs the ability of 53BP1 to bind the ubiquitinated H2A and to properly localize to the sites of DNA damage. Phosphorylation of 53BP1 at S1618 occurs at kinetochores and in cytosol and is restricted to mitotic cells. Interaction between 53BP1 and Plk1 depends on the activity of Cdk1. We propose that activity of Cdk1 and Plk1 allows spatiotemporally controlled suppression of 53BP1 function during mitosis.

Department of Cancer Cell Biology; Institute of Molecular Genetics; Academy of Sciences of the Czech Republic ; Prague Czech Republic

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Cell Cycle. 2015;14(9):1356-7. doi: 10.1080/15384101.2015.1024586. PubMed

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Jackson SP, Bartek J. The DNA-damage response in human biology and disease. Nature 2009; 461:1071-8; PMID:19847258; http://dx.doi.org/10.1038/nature08467 PubMed DOI PMC

Shiloh Y. ATM and related protein kinases: safeguarding genome integrity. Nat Rev Cancer 2003; 3:155-68; PMID:12612651; http://dx.doi.org/10.1038/nrc1011 PubMed DOI

Stucki M, Clapperton J, Mohammad D, Yaffe M, Smerdon S, Jackson S. MDC1 directly binds phosphorylated histone H2AX to regulate cellular responses to DNA double-strand breaks. Cell 2005; 123:1213-26; PMID: 16377563; http://dx.doi.org/10.1016/j.cell.2005.09.038 PubMed DOI

Mailand N, Bekker-Jensen S, Faustrup H, Melander F, Bartek J, Lukas C, Lukas J. RNF8 ubiquitylates histones at DNA double-strand breaks and promotes assembly of repair proteins. Cell 2007; 131:887-900; PMID:18001824; http://dx.doi.org/10.1016/j.cell.2007.09.040 PubMed DOI

Doil C, Mailand N, Bekker-Jensen S, Menard P, Larsen DH, Pepperkok R, Ellenberg J, Panier S, Durocher D, Bartek J, et al. . RNF168 binds and amplifies ubiquitin conjugates on damaged chromosomes to allow accumulation of repair proteins. Cell 2009; 136:435-46; PMID:19203579; http://dx.doi.org/10.1016/j.cell.2008.12.041 PubMed DOI

Mattiroli F, Vissers Joseph HA, van Dijk Willem J, Ikpa P, Citterio E, Vermeulen W, Marteijn Jurgen A, Sixma Titia K. RNF168 ubiquitinates K13-15 on H2A/H2AX to drive DNA damage signaling. Cell 2012; 150:1182-95; PMID:22980979; http://dx.doi.org/10.1016/j.cell.2012.08.005 PubMed DOI

Fradet-Turcotte A, Canny MD, Escribano-Diaz C, Orthwein A, Leung CCY, Huang H, Landry M-C, Kitevski-LeBlanc J, Noordermeer SM, Sicheri F, et al. . 53BP1 is a reader of the DNA-damage-induced H2A Lys 15 ubiquitin mark. Nature 2013; 499:50-4; PMID: 23760478; http://dx.doi.org/10.1038/nature12318 PubMed DOI PMC

Botuyan MV, Lee J, Ward IM, Kim J-E, Thompson JR, Chen J, Mer G. Structural basis for the methylation state-specific Recognition of Histone H4-K20 by 53BP1 and Crb2 in DNA Repair. Cell 2006; 127:1361-73; PMID:17190600; http://dx.doi.org/10.1016/j.cell.2006.10.043 PubMed DOI PMC

Wang B, Matsuoka S, Ballif BA, Zhang D, Smogorzewska A, Gygi SP, Elledge SJ. Abraxas and RAP80 form a BRCA1 protein complex required for the DNA damage response. Science 2007; 316:1194-8; PMID:17525340; http://dx.doi.org/10.1126/science.1139476 PubMed DOI PMC

Kim H, Chen J, Yu X. Ubiquitin-binding protein RAP80 mediates BRCA1-dependent DNA damage response. Science 2007; 316:1202-5; PMID:17525342; http://dx.doi.org/10.1126/science.1139621 PubMed DOI

Noon AT, Shibata A, Rief N, Lobrich M, Stewart GS, Jeggo PA, Goodarzi AA. 53BP1-dependent robust localized KAP-1 phosphorylation is essential for heterochromatic DNA double-strand break repair. Nat Cell Biol 2010; 12:177-84; PMID:20081839; http://dx.doi.org/10.1038/ncb2017 PubMed DOI

Escribano-Díaz C, Orthwein A, Fradet-Turcotte A, Xing M, Young Jordan TF, Tkáč J, Cook Michael A, Rosebrock Adam P, Munro M, Canny Marella D, et al. . A cell cycle-dependent regulatory circuit composed of 53BP1-RIF1 and BRCA1-CtIP controls DNA repair pathway choice. Mol Cell 2013; 49:872-83; http://dx.doi.org/10.1016/j.molcel.2013.01.001 PubMed DOI

Dimitrova N, Chen Y-CM, Spector DL, de Lange T. 53BP1 promotes non-homologous end joining of telomeres by increasing chromatin mobility. Nature 2008; 456:524-8; PMID:18931659; http://dx.doi.org/10.1038/nature07433 PubMed DOI PMC

Di Virgilio M, Callen E, Yamane A, Zhang W, Jankovic M, Gitlin AD, Feldhahn N, Resch W, Oliveira TY, Chait BT, et al. . Rif1 prevents resection of DNA breaks and promotes immunoglobulin class switching. Science 2013; 339:711-5; PMID:23306439; http://dx.doi.org/10.1126/science.1230624 PubMed DOI PMC

Chapman JR, Barral P, Vannier J-B, Borel V, Steger M, Tomas-Loba A, Sartori Alessandro A, Adams Ian R, Batista Facundo D, Boulton Simon J. RIF1 is essential for 53BP1-dependent nonhomologous end joining and suppression of DNA double-strand break resection. Mol Cell 2013; 49:858-71; PMID:23333305; http://dx.doi.org/10.1016/j.molcel.2013.01.002 PubMed DOI PMC

Zimmermann M, Lottersberger F, Buonomo SB, Sfeir A, de Lange T. 53BP1 regulates DSB repair using Rif1 to control 5′ end resection. Science 2013; 339:700-4; PMID: 23306437; http://dx.doi.org/10.1126/science.1231573 PubMed DOI PMC

Panier S, Boulton SJ. Double-strand break repair: 53BP1 comes into focus. Nat Rev Mol Cell Biol 2014; 15:7-18; PMID:24326623; http://dx.doi.org/10.1038/nrm3719 PubMed DOI

Zimmermann M, de Lange T. 53BP1: pro choice in DNA repair. Trends Cell Biol 2014; 24:108-17; PMID: 24094932; http://dx.doi.org/10.1016/j.tcb.2013.09.003 PubMed DOI PMC

Giunta S, Belotserkovskaya R, Jackson SP. DNA damage signaling in response to double-strand breaks during mitosis. J Cell Biol 2010; 190:197-207; PMID:20660628; http://dx.doi.org/10.1083/jcb.200911156 PubMed DOI PMC

Giunta S, Jackson SP. Give me a break, but not in mitosis: the mitotic DNA damage response marks DNA double-strand breaks with early signaling events. Cell Cycle 2011; 10:1215-21; PMID:21412056; http://dx.doi.org/10.4161/cc.10.8.15334 PubMed DOI PMC

Zhang W, Peng G, Lin S-Y, Zhang P. DNA damage response is suppressed by the high cyclin-dependent kinase 1 activity in mitotic mammalian cells. J Biol Chem 2011; 286:35899-905; PMID:21878640; http://dx.doi.org/10.1074/jbc.M111.267690 PubMed DOI PMC

Nelson G, Buhmann M, von Zglinicki T. DNA damage foci in mitosis are devoid of 53BP1. Cell Cycle 2009; 8:3379-83; PMID:19806024; http://dx.doi.org/10.4161/cc.8.20.9857 PubMed DOI

van Vugt MATM, Gardino AK, Linding R, Ostheimer GJ, Reinhardt HC, Ong S-E, Tan CS, Miao H, Keezer SM, Li J, et al. . A mitotic phosphorylation feedback network connects Cdk1, Plk1, 53BP1, and Chk2 to inactivate the G2/M DNA damage checkpoint. PLoS Biol 2010; 8:e1000287; PMID:20126263; http://dx.doi.org/10.1371/journal.pbio.1000287 PubMed DOI PMC

Macurek L, Benada J, Müllers E, Halim VA, Krejčíková K, Burdová K, Pecháčková S, Hodný Z, Lindqvist A, Medema RH, et al. . Downregulation of Wip1 phosphatase modulates the cellular threshold of DNA damage signaling in mitosis. Cell Cycle 2013; 12:251-62; PMID:23255129; http://dx.doi.org/10.4161/cc.23057 PubMed DOI PMC

Dephoure N, Zhou C, Villén J, Beausoleil SA, Bakalarski CE, Elledge SJ, Gygi SP. A quantitative atlas of mitotic phosphorylation. Proc Natl Acad Sci 2008; 105:10762-7; http://dx.doi.org/10.1073/pnas.0805139105 PubMed DOI PMC

Olsen JV, Vermeulen M, Santamaria A, Kumar C, Miller ML, Jensen LJ, Gnad F, Cox J, Jensen TS, Nigg EA, et al. . Quantitative phosphoproteomics reveals widespread full phosphorylation site occupancy during mitosis. Sci Signal 2010; 3:ra3-; PMID:20068231; http://dx.doi.org/10.1126/scisignal.2000475 PubMed DOI

Vassilev LT. Cell cycle synchronization at the G2/M phase border by reversible inhibition of CDK1. Cell Cycle 2006; 5:2555-6; PMID:17172841; http://dx.doi.org/10.4161/cc.5.22.3463 PubMed DOI

Vassilev LT, Tovar C, Chen S, Knezevic D, Zhao X, Sun H, Heimbrook DC, Chen L. Selective small-molecule inhibitor reveals critical mitotic functions of human CDK1. Proc Natl Acad Sci 2006; 103:10660-5; http://dx.doi.org/10.1073/pnas.0600447103 PubMed DOI PMC

Seki A, Coppinger JA, Du H, Jang C-Y, Yates JR, Fang G. Plk1- and β-TrCP–dependent degradation of Bora controls mitotic progression. J Cell Biol 2008; 181:65-78; PMID:18378770; http://dx.doi.org/10.1083/jcb.200712027 PubMed DOI PMC

Bruinsma W, Raaijmakers JA, Medema RH. Switching Polo-like kinase-1 on and off in time and space. Trends Biochem Sci 2012; 37:534-42; PMID:23141205; http://dx.doi.org/10.1016/j.tibs.2012.09.005 PubMed DOI

Takaki T, Trenz K, Costanzo V, Petronczki M. Polo-like kinase 1 reaches beyond mitosis—cytokinesis, DNA damage response, and development. Curr Opin Cell Biol 2008; 20:650-60; PMID:19000759; http://dx.doi.org/10.1016/j.ceb.2008.10.005 PubMed DOI

van Vugt MATM, Brás A, Medema RH. Polo-like kinase-1 controls recovery from a G2 DNA damage-induced arrest in mammalian cells. Mol Cell 2004; 15:799-811; PMID:15350223; http://dx.doi.org/10.1016/j.molcel.2004.07.015 PubMed DOI

Medema RH, Macurek L. Checkpoint control and cancer. Oncogene 2012; 31:2601-13; PMID:21963855; http://dx.doi.org/10.1038/onc.2011.451 PubMed DOI

Mamely I, van Vugt MATM, Smits VAJ, Semple JI, Lemmens B, Perrakis A, Medema RH, Freire R. Polo-like kinase-1 controls proteasome-dependent degradation of claspin during checkpoint recovery. Curr Biol 2006; 16:1950-5; PMID:16934469; http://dx.doi.org/10.1016/j.cub.2006.08.026 PubMed DOI

Mailand N, Bekker-Jensen S, Bartek J, Lukas J. Destruction of claspin by SCF[β]TrCP restrains Chk1 activation and facilitates recovery from genotoxic stress. Mol Cell 2006; 23:307-18; PMID:16885021; http://dx.doi.org/10.1016/j.molcel.2006.06.016 PubMed DOI

Nakajima H, Toyoshima-Morimoto F, Taniguchi E, Nishida E. Identification of a consensus motif for Plk (Polo-like Kinase) phosphorylation reveals Myt1 as a Plk1 substrate*. J Biol Chem 2003; 278:25277-80; PMID:12738781; http://dx.doi.org/10.1074/jbc.C300126200 PubMed DOI

Santamaria A, Wang B, Elowe S, Malik R, Zhang F, Bauer M, Schmidt A, Silljé HHW, Körner R, Nigg EA. The Plk1-dependent phosphoproteome of the early mitotic spindle. Mol Cell Proteomics 2011; 10:M110.004457; PMID:20860994; http://dx.doi.org/10.1074/mcp.M110.004457 PubMed DOI PMC

Heijink A, Krajewska M, van Vugt MA. The DNA damage response during mitosis. Mutat Res 2013; 750:45-55; PMID:23880065; http://dx.doi.org/10.1016/j.mrfmmm.2013.07.003 PubMed DOI

Petronczki M, Lénárt P, Peters J-M. Polo on the rise—from mitotic entry to cytokinesis with Plk1. Dev Cell 2008; 14:646-59; PMID:18477449; http://dx.doi.org/10.1016/j.devcel.2008.04.014 PubMed DOI

Jullien D, Vagnarelli P, Earnshaw WC, Adachi Y. Kinetochore localisation of the DNA damage response component 53BP1 during mitosis. J Cell Sci 2002; 115:71-9; PMID:11801725 PubMed

Lindon C, Pines J. Ordered proteolysis in anaphase inactivates Plk1 to contribute to proper mitotic exit in human cells. J Cell Biol 2004; 164:233-41; PMID:14734534; http://dx.doi.org/10.1083/jcb.200309035 PubMed DOI PMC

Cha H, Wang X, Li H, Fornace AJ. A functional role for p38 MAPK in modulating mitotic transit in the absence of stress. J Biol Chem 2007; 282:22984-92; PMID:17548358; http://dx.doi.org/10.1074/jbc.M700735200 PubMed DOI

Lee K, Kenny AE, Rieder CL. P38 mitogen-activated protein kinase activity is required during mitosis for timely satisfaction of the mitotic checkpoint but not for the fidelity of chromosome segregation. Mol Biol Cell 2010; 21:2150-60; PMID:20462950; http://dx.doi.org/10.1091/mbc.E10-02-0125 PubMed DOI PMC

Elia AEH, Rellos P, Haire LF, Chao JW, Ivins FJ, Hoepker K, Mohammad D, Cantley LC, Smerdon SJ, Yaffe MB. The molecular basis for phosphodependent substrate targeting and regulation of Plks by the polo-box domain. Cell 2003; 115:83-95; PMID:14532005; http://dx.doi.org/10.1016/S0092-8674(03)00725-6 PubMed DOI

Gatti M, Pinato S, Maspero E, Soffientini P, Polo S, Penengo L. A novel ubiquitin mark at the N-terminal tail of histone H2As targeted by RNF168 ubiquitin ligase. Cell Cycle 2012; 11:2538-44; PMID:22713238; http://dx.doi.org/10.4161/cc.20919 PubMed DOI PMC

Harrigan JA, Belotserkovskaya R, Coates J, Dimitrova DS, Polo SE, Bradshaw CR, Fraser P, Jackson SP. Replication stress induces 53BP1-containing OPT domains in G1 cells. J Cell Biol 2011; 193:97-108; PMID:21444690 PubMed PMC

Lukas C, Savic V, Bekker-Jensen S, Doil C, Neumann B, Solvhoj Pedersen R, Grofte M, Chan KL, Hickson ID, Bartek J, et al. . 53BP1 nuclear bodies form around DNA lesions generated by mitotic transmission of chromosomes under replication stress. Nat Cell Biol 2011; 13:243-53; PMID:21317883; http://dx.doi.org/10.1038/ncb2201 PubMed DOI

Macurek L, Lindqvist A, Lim D, Lampson MA, Klompmaker R, Freire R, Clouin C, Taylor SS, Yaffe MB, Medema RH. Polo-like kinase-1 is activated by aurora a to promote checkpoint recovery. Nature 2008; 455:119-23; PMID:18615013; http://dx.doi.org/10.1038/nature07185 PubMed DOI

Hori T, Shang W-H, Toyoda A, Misu S, Monma N, Ikeo K, Molina O, Vargiu G, Fujiyama A, Kimura H, et al. . Histone H4 Lys 20 monomethylation of the CENP-A nucleosome is essential for kinetochore assembly. Dev Cell 2014; 29:740-9; PMID:24960696; http://dx.doi.org/10.1016/j.devcel.2014.05.001 PubMed DOI PMC

Lee D-H, Acharya Sanket S, Kwon M, Drane P, Guan Y, Adelmant G, Kalev P, Shah J, Pellman D, Marto Jarrod A, et al. . Dephosphorylation enables the recruitment of 53BP1 to double-strand DNA breaks. Mol Cell 2014; 54:512-25; PMID:24703952; http://dx.doi.org/10.1016/j.molcel.2014.03.020 PubMed DOI PMC

Orthwein A, Fradet-Turcotte A, Noordermeer SM, Canny MD, Brun CM, Strecker J, Escribano-Diaz C, Durocher D. Mitosis inhibits DNA double-strand break repair to guard against telomere fusions. Science 2014; 344:189-93; PMID:24652939; http://dx.doi.org/10.1126/science.1248024 PubMed DOI

Sharma N, Zhu Q, Wani G, He J, Wang Q-e, Wani AA. USP3 counteracts RNF168 via deubiquitinating H2A and γH2AX at lysine 13 and 15. Cell Cycle 2014; 13:106-14; PMID:24196443; http://dx.doi.org/10.4161/cc.26814 PubMed DOI PMC

Mosbech A, Lukas C, Bekker-Jensen S, Mailand N. The deubiquitylating enzyme USP44 counteracts the DNA double-strand break response mediated by the RNF8 and RNF168 ubiquitin ligases. J Biol Chem 2013; 288:16579-87; PMID:23615962; http://dx.doi.org/10.1074/jbc.M113.459917 PubMed DOI PMC

Delgado-Díaz MR, Martín Y, Berg A, Freire R, Smits VAJ. Dub3 controls DNA damage signalling by direct deubiquitination of H2AX. Mol Oncol 2014; 8(5):884-93; PMID:24704006 PubMed PMC

Joo H-Y, Zhai L, Yang C, Nie S, Erdjument-Bromage H, Tempst P, Chang C, Wang H. Regulation of cell cycle progression and gene expression by H2A deubiquitination. Nature 2007; 449:1068-72; PMID:17914355; http://dx.doi.org/10.1038/nature06256 PubMed DOI

Xu Y, Yang H, Joo H-Y, Yu J-H, Smith Iv AD, Schneider D, Chow LT, Renfrow M, Wang H. Ubp-M serine 552 phosphorylation by cyclin-dependent kinase 1 regulates cell cycle progression. Cell Cycle 2013; 12:3408-16 PubMed PMC

Cai S-Y, Babbitt RW, Marchesi VT. A mutant deubiquitinating enzyme (Ubp-M) associates with mitotic chromosomes and blocks cell division. Proc Natl Acad Sci 1999; 96:2828-33; http://dx.doi.org/10.1073/pnas.96.6.2828 PubMed DOI PMC

Simoneau A, Robellet X, Ladouceur A-M, D’Amours D. Cdk1-dependent regulation of the Mre11 complex couples DNA repair pathways to cell cycle progression. Cell Cycle 2014; 13:1078-90; PMID:24553123; http://dx.doi.org/10.4161/cc.27946 PubMed DOI PMC

Esashi F, Christ N, Gannon J, Liu Y, Hunt T, Jasin M, West SC. CDK-dependent phosphorylation of BRCA2 as a regulatory mechanism for recombinational repair. Nature 2005; 434:598-604; PMID:15800615; http://dx.doi.org/10.1038/nature03404 PubMed DOI

Hayashi MT, Cesare AJ, Fitzpatrick JAJ, Lazzerini-Denchi E, Karlseder J. A telomere-dependent DNA damage checkpoint induced by prolonged mitotic arrest. Nat Struct Mol Biol 2012; 19:387-94; PMID:22407014; http://dx.doi.org/10.1038/nsmb.2245 PubMed DOI PMC

Knobel PA, Belotserkovskaya R, Galanty Y, Schmidt CK, Jackson SP, Stracker TH. USP28 is recruited to sites of DNA damage by the tandem BRCT domains of 53BP1 but plays a minor role in double-strand break metabolism. Mol Cell Biol 2014; 34:2062-74; PMID:24687851; http://dx.doi.org/10.1128/MCB.00197-14 PubMed DOI PMC

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