title: Direct involvement of retinoblastoma family proteins in DNA repair by non-homologous end-joining.
abstract: Deficiencies in DNA double-strand break (DSB) repair lead to genetic instability, a recognized cause of cancer initiation and evolution. We report that the retinoblastoma tumor suppressor protein (RB1) is required for DNA DSB repair by canonical non-homologous end-joining (cNHEJ). Support of cNHEJ involves a mechanism independent of RB1's cell-cycle function and depends on its amino terminal domain with which it binds to NHEJ components XRCC5 and XRCC6. Cells with engineered loss of RB family function as well as cancer-derived cells with mutational RB1 loss show substantially reduced levels of cNHEJ. RB1 variants disabled for the interaction with XRCC5 and XRCC6, including a cancer-associated variant, are unable to support cNHEJ despite being able to confer cell-cycle control. Our data identify RB1 loss as a candidate driver of structural genomic instability and a causative factor for cancer somatic heterogeneity and evolution.
authors.affiliation: Division of Cancer Biology, Institute of Cancer Research, 237 Fulham Road, London SW3 6JB, UK; Cancer Cell Signalling, UCL Cancer Institute, University College London, 72 Huntley Street, London WC1E 6DD, UK.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Cook
authors.affiliation: Cancer Cell Signalling, UCL Cancer Institute, University College London, 72 Huntley Street, London WC1E 6DD, UK.
authors.forename: Georgia
authors.initials: G
authors.lastname: Zoumpoulidou
authors.affiliation: Division of Cancer Biology, Institute of Cancer Research, 237 Fulham Road, London SW3 6JB, UK.
authors.forename: Maciej T
authors.initials: MT
authors.lastname: Luczynski
authors.affiliation: Cancer Cell Signalling, UCL Cancer Institute, University College London, 72 Huntley Street, London WC1E 6DD, UK.
authors.forename: Simone
authors.initials: S
authors.lastname: Rieger
authors.affiliation: Public Health England, Centre for Radiation, Chemical and Environmental Hazards, Chilton, Didcot OX11 0RQ, UK.
authors.forename: Jayne
authors.initials: J
authors.lastname: Moquet
authors.affiliation: Cancer Research UK Drug-DNA Interactions Research Group, UCL Cancer Institute, University College London, 72 Huntley Street, London WC1E 6DD, UK.
authors.forename: Victoria J
authors.initials: VJ
authors.lastname: Spanswick
authors.affiliation: Cancer Research UK Drug-DNA Interactions Research Group, UCL Cancer Institute, University College London, 72 Huntley Street, London WC1E 6DD, UK.
authors.forename: John A
authors.initials: JA
authors.lastname: Hartley
authors.affiliation: Public Health England, Centre for Radiation, Chemical and Environmental Hazards, Chilton, Didcot OX11 0RQ, UK.
authors.forename: Kai
authors.initials: K
authors.lastname: Rothkamm
authors.affiliation: Division of Cancer Biology, Institute of Cancer Research, 237 Fulham Road, London SW3 6JB, UK. Electronic address: paul.huang@icr.ac.uk.
authors.forename: Paul H
authors.initials: PH
authors.lastname: Huang
authors.affiliation: Division of Cancer Biology, Institute of Cancer Research, 237 Fulham Road, London SW3 6JB, UK; Cancer Cell Signalling, UCL Cancer Institute, University College London, 72 Huntley Street, London WC1E 6DD, UK. Electronic address: s.mittnacht@ucl.ac.uk.
authors.forename: Sibylle
authors.initials: S
authors.lastname: Mittnacht
mesh_terms: D034961:Antigens, Nuclear / Q000378:metabolism*; D002453:Cell Cycle / Q000502:physiology; D053903:DNA Breaks, Double-Stranded; D059766:DNA End-Joining Repair / Q000235:genetics*; D004265:DNA Helicases / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D042822:Genomic Instability / Q000235:genetics*; D006801:Humans; D000072200:Ku Autoantigen; D011995:Recombination, Genetic / Q000235:genetics; D016160:Retinoblastoma Protein / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000378:metabolism
chemical_list: D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D016160:Retinoblastoma Protein; D025521:Tumor Suppressor Proteins; D004265:DNA Helicases; C106893:XRCC5 protein, human; C000606509:Xrcc6 protein, human; D000072200:Ku Autoantigen
references.citation: Nat Rev Mol Cell Biol. 2008 Apr;9(4):297-308
references.pmid: 18285803
references.citation: Radiat Oncol. 2011;6:106
references.pmid: 21888676
references.citation: Cell. 1992 Sep 18;70(6):993-1006
references.pmid: 1388095
references.citation: J Radiat Res. 2010;51(6):633-41
references.pmid: 21116096
references.citation: Cancer Res. 2006 May 15;66(10):5354-62
references.pmid: 16707462
references.citation: DNA Repair (Amst). 2014 Apr;16:84-96
references.pmid: 24629483
references.citation: Am J Hum Genet. 1997 Aug;61(2):282-94
references.pmid: 9311732
references.citation: Immunol Res. 2012 Dec;54(1-3):233-46
references.pmid: 22569912
references.citation: DNA Repair (Amst). 2014 Jul;19:169-75
references.pmid: 24746645
references.citation: PLoS Genet. 2008 Jun;4(6):e1000110
references.pmid: 18584027
references.citation: Oncogene. 1998 Oct 29;17(17):2177-86
references.pmid: 9811449
references.citation: Cell Cycle. 2014;13(19):3016-25
references.pmid: 25486562
references.citation: Annu Rev Biochem. 2010;79:181-211
references.pmid: 20192759
references.citation: Nat Rev Mol Cell Biol. 2014 Jan;15(1):7-18
references.pmid: 24326623
references.citation: J Biol Chem. 1994 Jan 14;269(2):1380-7
references.pmid: 8288605
references.citation: Clin Experiment Ophthalmol. 2014 Jan-Feb;42(1):33-52
references.pmid: 24433356
references.citation: Expert Rev Proteomics. 2010 Jun;7(3):401-9
references.pmid: 20536310
references.citation: Acta Oncol. 2008;47(8):1483-92
references.pmid: 18785023
references.citation: Mol Cell. 2007 Nov 9;28(3):371-85
references.pmid: 17996702
references.citation: Nat Rev Mol Cell Biol. 2003 Sep;4(9):712-20
references.pmid: 14506474
references.citation: Cancer Res. 2013 Jul 15;73(14):4247-55
references.pmid: 23687339
references.citation: Cell Div. 2012 Mar 14;7(1):10
references.pmid: 22417103
references.citation: Cell Rep. 2014 Jul 24;8(2):430-8
references.pmid: 25001286
references.citation: Nat Methods. 2013 Aug;10(8):730-6
references.pmid: 23921808
references.citation: DNA Repair (Amst). 2006 Sep 8;5(9-10):1042-8
references.pmid: 16822724
references.citation: Adv Cancer Res. 2014;121:147-82
references.pmid: 24889531
references.citation: Cold Spring Harb Perspect Biol. 2014 Dec;6(12):a016576
references.pmid: 25228584
references.citation: J Biol Chem. 1997 May 9;272(19):12738-46
references.pmid: 9139732
references.citation: Nat Struct Mol Biol. 2010 Sep;17(9):1144-51
references.pmid: 20802485
references.citation: Genes Dev. 2002 Dec 15;16(24):3199-212
references.pmid: 12502741
references.citation: J Clin Oncol. 2012 Mar 20;30(9):950-7
references.pmid: 22355046
references.citation: J Clin Oncol. 2009 May 10;27(14):2356-62
references.pmid: 19255307
references.citation: J Cell Physiol. 2003 Nov;197(2):169-80
references.pmid: 14502556
references.citation: Mutagenesis. 2014 Mar;29(2):123-9
references.pmid: 24452505
references.citation: Proc Natl Acad Sci U S A. 2006 Jun 13;103(24):9057-62
references.pmid: 16754850
references.citation: Genome Biol. 2003;4(5):P3
references.pmid: 12734009
references.citation: Blood. 2008 Feb 15;111(4):1894-902
references.pmid: 18048646
references.citation: Oncol Res. 1995;7(2):63-6
references.pmid: 7579728
references.citation: J Clin Oncol. 2014 Oct 10;32(29):3284-90
references.pmid: 25185089
references.citation: Genes Dev. 2013 May 1;27(9):1003-15
references.pmid: 23618872
references.citation: In Vivo. 2008 May-Jun;22(3):305-9
references.pmid: 18610740
references.citation: Cell Stem Cell. 2008 Oct 9;3(4):416-28
references.pmid: 18940733
references.citation: Nat Rev Cancer. 2012 Mar;12(3):220-6
references.pmid: 22318235
references.citation: Nat Rev Genet. 2013 Jan;14(1):62-75
references.pmid: 23247436
references.citation: Mol Cell Biol. 2003 Aug;23(16):5706-15
references.pmid: 12897142
references.citation: Genes Dev. 2000 Dec 1;14(23):3051-64
references.pmid: 11114893
references.citation: Nature. 1992 Sep 24;359(6393):288-94
references.pmid: 1406932
references.citation: Int J Mol Med. 2013 Jul;32(1):137-43
references.pmid: 23670186
references.citation: Nat Rev Mol Cell Biol. 2010 Mar;11(3):220-8
references.pmid: 20177397
references.citation: PLoS One. 2012;7(2):e31627
references.pmid: 22384045
references.citation: Cytogenet Genome Res. 2004;104(1-4):14-20
references.pmid: 15162010
references.citation: Nature. 1998 Feb 26;391(6670):859-65
references.pmid: 9495340
references.citation: Hum Mol Genet. 1995 Mar;4(3):383-8
references.pmid: 7795591
references.citation: Proc Natl Acad Sci U S A. 2003 Mar 4;100(5):2363-8
references.pmid: 12598654
references.citation: Hum Mol Genet. 2008 May 15;17(10):1363-72
references.pmid: 18211953
references.citation: Annu Rev Genet. 2010;44:113-39
references.pmid: 20690856
references.citation: Mol Cell. 2012 Aug 24;47(4):497-510
references.pmid: 22920291
references.citation: Ann N Y Acad Sci. 2012 Sep;1267:110-6
references.pmid: 22954224
references.citation: Adv Exp Med Biol. 2010;685:220-7
references.pmid: 20687510
references.citation: Br J Radiol. 2014 Mar;87(1035):20130685
references.pmid: 24363387
references.citation: Mol Cancer Ther. 2004 Nov;3(11):1427-38
references.pmid: 15542782
references.citation: Nature. 1992 Sep 24;359(6393):295-300
references.pmid: 1406933
references.citation: Cold Spring Harb Perspect Biol. 2013 May;5(5):a012757
references.pmid: 23637284
references.citation: Proc Natl Acad Sci U S A. 2014 Aug 5;111(31):11341-6
references.pmid: 25049398
references.citation: Cell Cycle. 2012 Sep 1;11(17):3189-98
references.pmid: 22895179
references.citation: J Gene Med. 2005 Apr;7(4):407-19
references.pmid: 15538723