Phospho-Ku70 induced by DNA damage interacts with RNA Pol II and promotes the formation of phospho-53BP1 foci to ensure optimal cNHEJ.
Journal
Nucleic acids research
ISSN: 1362-4962
Titre abrégé: Nucleic Acids Res
Pays: England
ID NLM: 0411011
Informations de publication
Date de publication:
18 11 2021
18 11 2021
Historique:
accepted:
26
10
2021
revised:
15
09
2021
received:
23
04
2021
pubmed:
1
11
2021
medline:
28
12
2021
entrez:
31
10
2021
Statut:
ppublish
Résumé
Canonical non-homologous end-joining (cNHEJ) is the prominent mammalian DNA double-strand breaks (DSBs) repair pathway operative throughout the cell cycle. Phosphorylation of Ku70 at ser27-ser33 (pKu70) is induced by DNA DSBs and has been shown to regulate cNHEJ activity, but the underlying mechanism remained unknown. Here, we established that following DNA damage induction, Ku70 moves from nucleoli to the sites of damage, and once linked to DNA, it is phosphorylated. Notably, the novel emanating functions of pKu70 are evidenced through the recruitment of RNA Pol II and concomitant formation of phospho-53BP1 foci. Phosphorylation is also a prerequisite for the dynamic release of Ku70 from the repair complex through neddylation-dependent ubiquitylation. Although the non-phosphorylable ala-Ku70 form does not compromise the formation of the NHEJ core complex per se, cells expressing this form displayed constitutive and stress-inducible chromosomal instability. Consistently, upon targeted induction of DSBs by the I-SceI meganuclease into an intrachromosomal reporter substrate, cells expressing pKu70, rather than ala-Ku70, are protected against the joining of distal DNA ends. Collectively, our results underpin the essential role of pKu70 in the orchestration of DNA repair execution in living cells and substantiated the way it paves the maintenance of genome stability.
Identifiants
pubmed: 34718776
pii: 6414600
doi: 10.1093/nar/gkab980
pmc: PMC8599715
doi:
Substances chimiques
Tumor Suppressor p53-Binding Protein 1
0
RNA Polymerase II
EC 2.7.7.-
Ku Autoantigen
EC 4.2.99.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
11728-11745Informations de copyright
© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.
Références
Genes Dev. 2001 Jan 1;15(1):50-65
pubmed: 11156605
Proc Natl Acad Sci U S A. 2020 Mar 10;117(10):5329-5338
pubmed: 32094185
Nucleic Acids Res. 2016 Sep 19;44(16):7755-65
pubmed: 27402161
Blood. 2010 Jul 15;116(2):239-49
pubmed: 20424183
Breast Cancer Res. 2017 Oct 18;19(1):115
pubmed: 29047390
Genes Dev. 1999 Oct 15;13(20):2633-8
pubmed: 10541549
Nat Commun. 2016 Dec 06;7:13785
pubmed: 27922005
Nucleic Acids Res. 2002 Aug 1;30(15):3454-63
pubmed: 12140331
Nucleic Acids Res. 2020 Nov 4;48(19):10953-10972
pubmed: 33045735
Blood. 2005 Jun 15;105(12):4776-83
pubmed: 15718417
J Cell Biol. 2013 Aug 5;202(3):579-95
pubmed: 23897892
Mol Cell. 2017 Jun 15;66(6):801-817
pubmed: 28622525
Nat Genet. 1999 Oct;23(2):194-8
pubmed: 10508516
Nat Struct Mol Biol. 2009 Aug;16(8):814-8
pubmed: 19633669
Nat Rev Cancer. 2016 Jan;16(1):35-42
pubmed: 26667849
Nat Commun. 2017 Dec 11;8(1):2039
pubmed: 29229926
Oncotarget. 2015 Sep 29;6(29):27980-8000
pubmed: 26337656
Cell. 2021 Mar 4;184(5):1314-1329.e10
pubmed: 33626331
Cell. 2006 Dec 29;127(7):1361-73
pubmed: 17190600
Science. 1989 Nov 3;246(4930):629-34
pubmed: 2683079
J Cell Sci. 2015 Feb 15;128(4):607-20
pubmed: 25609713
Mol Cell. 2016 Jan 7;61(1):15-26
pubmed: 26687679
Cell. 2016 Nov 3;167(4):1001-1013.e7
pubmed: 27881299
Nature. 2001 Aug 9;412(6847):607-14
pubmed: 11493912
Genes Dev. 2011 Mar 1;25(5):409-33
pubmed: 21363960
Nature. 2018 Aug;560(7716):122-127
pubmed: 30046110
Nat Rev Mol Cell Biol. 2017 Aug;18(8):495-506
pubmed: 28512351
Nat Struct Mol Biol. 2009 Aug;16(8):808-13
pubmed: 19633670
Nat Commun. 2016 Apr 11;7:11242
pubmed: 27063109
J Cell Sci. 2004 Aug 1;117(Pt 17):3935-45
pubmed: 15265990
Nat Cell Biol. 2017 Dec;19(12):1400-1411
pubmed: 29180822
Cell Rep. 2013 Aug 29;4(4):817-29
pubmed: 23972994
Mutat Res. 2011 Jun 3;711(1-2):61-72
pubmed: 21329706
Mol Cell Oncol. 2014 Oct 30;2(1):e968020
pubmed: 27308383
Cell Rep. 2017 Jan 10;18(2):520-532
pubmed: 28076794
Nat Commun. 2016 Oct 05;7:13049
pubmed: 27703167
PLoS Genet. 2016 Oct 31;12(10):e1006230
pubmed: 27798638
Nat Cell Biol. 2019 Oct;21(10):1286-1299
pubmed: 31570834
Genome Integr. 2012 Nov 27;3(1):9
pubmed: 23181949
Mol Cell. 2004 Jun 4;14(5):611-23
pubmed: 15175156
Cell Rep. 2015 May 5;11(5):704-14
pubmed: 25921528
Annu Rev Biophys. 2015;44:207-28
pubmed: 26098514
Nat Commun. 2020 Oct 16;11(1):5239
pubmed: 33067475
Cancer Res. 2016 Sep 15;76(18):5189-91
pubmed: 27635040
J Exp Clin Cancer Res. 2016 Sep 27;35(1):153
pubmed: 27670139
DNA Repair (Amst). 2014 May;17:21-9
pubmed: 24680878
Mutat Res. 2010 Apr-Jun;704(1-3):2-11
pubmed: 20006736
Nature. 2017 Sep 20;549(7673):548-552
pubmed: 28959974
Mol Cell. 2016 Oct 6;64(1):189-198
pubmed: 27716483
Nat Struct Mol Biol. 2009 Aug;16(8):819-24
pubmed: 19633668
J Biol Chem. 2009 Apr 10;284(15):9845-53
pubmed: 19176521
Mol Cell. 2010 Oct 22;40(2):179-204
pubmed: 20965415
PLoS Genet. 2014 Jan;10(1):e1004086
pubmed: 24453986
Nat Rev Mol Cell Biol. 2014 Jan;15(1):7-18
pubmed: 24326623
Nat Protoc. 2006;1(6):3129-42
pubmed: 17406576
Nucleic Acids Res. 2016 Feb 29;44(4):1732-45
pubmed: 26712563
Mol Cell Biol. 2011 Nov;31(21):4379-89
pubmed: 21876003
Mol Cell. 2017 Feb 16;65(4):671-684.e5
pubmed: 28132842
Environ Mol Mutagen. 2017 Jun;58(5):235-263
pubmed: 28485537
EMBO Rep. 2019 May;20(5):
pubmed: 30948458
Nat Struct Mol Biol. 2010 Jun;17(6):688-95
pubmed: 20453858
Mol Biol Cell. 2003 Oct;14(10):4221-9
pubmed: 12937274
Mol Cell. 2004 Dec 3;16(5):715-24
pubmed: 15574327
J Cell Sci. 2012 Aug 1;125(Pt 15):3529-34
pubmed: 22553214
J Cell Sci. 2017 Oct 1;130(19):3347-3359
pubmed: 28751496
Biochem J. 2015 Oct 1;471(1):1-11
pubmed: 26392571
Genome Biol. 2006;7(10):R100
pubmed: 17076895
Nature. 2019 Oct;574(7779):571-574
pubmed: 31645724
Biochem Biophys Res Commun. 2013 May 31;435(2):260-6
pubmed: 23639616
Cell. 1993 Jan 15;72(1):131-42
pubmed: 8422676
Annu Rev Genet. 2013;47:433-55
pubmed: 24050180