title: Remodeling and spacing factor 1 (RSF1) deposits centromere proteins at DNA double-strand breaks to promote non-homologous end-joining.
abstract: The cellular response to ionizing radiation (IR)-induced DNA double-strand breaks (DSBs) in native chromatin requires a tight coordination between the activities of DNA repair machineries and factors that modulate chromatin structure. SMARCA5 is an ATPase of the SNF2 family of chromatin remodeling factors that has recently been implicated in the DSB response. It forms distinct chromatin remodeling complexes with several non-canonical subunits, including the remodeling and spacing factor 1 (RSF1) protein. Despite the fact that RSF1 is often overexpressed in tumors and linked to tumorigenesis and genome instability, its role in the DSB response remains largely unclear. Here we show that RSF1 accumulates at DSB sites and protects human cells against IR-induced DSBs by promoting repair of these lesions through homologous recombination (HR) and non-homologous end-joining (NHEJ). Although SMARCA5 regulates the RNF168-dependent ubiquitin response that targets BRCA1 to DSBs, we found RSF1 to be dispensable for this process. Conversely, we found that RSF1 facilitates the assembly of centromere proteins CENP-S and CENP-X at sites of DNA damage, while SMARCA5 was not required for these events. Mechanistically, we uncovered that CENP-S and CENP-X, upon their incorporation by RSF1, promote assembly of the NHEJ factor XRCC4 at damaged chromatin. In contrast, CENP-S and CENP-X were dispensable for HR, suggesting that RSF1 regulates HR independently of these centromere proteins. Our findings reveal distinct functions of RSF1 in the 2 major pathways of DSB repair and explain how RSF1, through the loading of centromere proteins and XRCC4 at DSBs, promotes repair by non-homologous end-joining.
authors: Department of Toxicogenetics; Leiden University Medical Center; Leiden, The Netherlands.
authors: Angela
authors: A
authors: Helfricht
authors: Wouter W
authors: WW
authors: Wiegant
authors: Peter E
authors: PE
authors: Thijssen
authors: Alfred C
authors: AC
authors: Vertegaal
authors: Martijn S
authors: MS
authors: Luijsterburg
authors: Haico
authors: H
authors: van Attikum
mesh_terms: D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D051017:Apoptosis Regulatory Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D019313:BRCA1 Protein / Q000378:metabolism; D045744:Cell Line, Tumor; D002868:Chromosomal Proteins, Non-Histone / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D053903:DNA Breaks, Double-Stranded / Q000528:radiation effects; D059766:DNA End-Joining Repair; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D059765:Homologous Recombination; D006801:Humans; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D011839:Radiation, Ionizing; D015534:Trans-Activators / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D025801:Ubiquitin / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism
chemical_list: D051017:Apoptosis Regulatory Proteins; D019313:BRCA1 Protein; C493243:CENPS protein, human; C551977:CENPX protein, human; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D034741:RNA, Small Interfering; C452851:RSF1 protein, human; D015534:Trans-Activators; D025521:Tumor Suppressor Proteins; D025801:Ubiquitin; C101467:XRCC4 protein, human; C535210:RNF168 protein, human; D044767:Ubiquitin-Protein Ligases; D000251:Adenosine Triphosphatases; C409462:SMARCA5 protein, human
keywords: DNA double-strand break repair; RSF1; SMARCA5/SNF2H; centromere proteins; chromatin remodeling; non-homologous end-joining
references: Mol Oncol. 2011 Aug;5(4):387-93
references: 21821475
references: J Cell Biol. 2009 Jul 27;186(2):173-82
references: 19620631
references: Nature. 2007 Nov 22;450(7169):509-14
references: 17965729
references: J Cell Biol. 2009 May 4;185(3):397-407
references: 19398759
references: Nature. 2009 Oct 22;461(7267):1071-8
references: 19847258
references: Nature. 2013 Jan 17;493(7432):356-63
references: 23325218
references: Cell. 2007 Nov 30;131(5):887-900
references: 18001824
references: Mol Cell. 2011 Mar 4;41(5):515-28
references: 21362548
references: Nat Cell Biol. 2006 May;8(5):458-69
references: 16622419
references: Cell. 2009 Feb 6;136(3):435-46
references: 19203579
references: Annu Rev Biochem. 2013;82:55-80
references: 23414304
references: Mol Cell. 2010 Mar 26;37(6):879-86
references: 20347429
references: J Cell Biol. 2010 Sep 6;190(5):731-40
references: 20805324
references: Cell. 2009 Feb 6;136(3):420-34
references: 19203578
references: Mol Cell. 2010 Dec 22;40(6):976-87
references: 21172662
references: Nature. 2012 Sep 27;489(7417):581-4
references: 22960744
references: PLoS Genet. 2008 Jun 27;4(6):e1000110
references: 18584027
references: J Biol Chem. 2010 Dec 3;285(49):38260-9
references: 20923775
references: Nat Struct Mol Biol. 2011 Nov 27;18(12):1345-50
references: 22120668
references: Nucleic Acids Res. 2011 Oct;39(19):8445-56
references: 21745822
references: Cell Death Differ. 2013 Mar;20(3):490-502
references: 23197296
references: Mol Cell. 2010 Mar 26;37(6):865-78
references: 20347428
references: Cell. 2010 Jun 11;141(6):970-81
references: 20550933
references: J Cell Sci. 2013 Feb 15;126(Pt 4):889-903
references: 23264744
references: Proc Natl Acad Sci U S A. 2009 Sep 15;106(37):15762-7
references: 19717431
references: EMBO J. 2012 May 30;31(11):2511-27
references: 22531782
references: Methods Enzymol. 2006;409:524-40
references: 16793422
references: J Cell Biol. 2012 Apr 16;197(2):267-81
references: 22492724
references: Cell. 2007 Nov 30;131(5):901-14
references: 18001825
references: Cell. 2012 Feb 3;148(3):487-501
references: 22304917
references: Genes Dev. 1999 Oct 15;13(20):2633-8
references: 10541549
references: Trends Mol Med. 2007 Sep;13(9):373-80
references: 17822959
references: Mol Cell. 2012 Aug 24;47(4):497-510
references: 22920291
references: J Cell Sci. 2007 Aug 1;120(Pt 15):2731-40
references: 17646676
references: Mol Oncol. 2011 Aug;5(4):349-67
references: 21782533
references: Proc Natl Acad Sci U S A. 2005 Sep 27;102(39):14004-9
references: 16172393
references: Nat Commun. 2012 Apr 17;3:782
references: 22510687
references: Nature. 2011 Jun 08;474(7350):230-4
references: 21654808
references: Nucleic Acids Res. 2013 Mar 1;41(5):2950-62
references: 23358825
references: Proc Natl Acad Sci U S A. 2007 Dec 26;104(52):20759-63
references: 18077395
references: Mol Cell. 2010 Oct 22;40(2):179-204
references: 20965415
references: J Cell Biol. 2010 Sep 6;190(5):741-9
references: 20805320
references: EMBO J. 2010 Sep 15;29(18):3130-9
references: 20693977
references: Hum Mol Genet. 2004 Oct 15;13(20):2369-76
references: 15333585
references: Science. 2008 Jun 13;320(5882):1507-10
references: 18483401