title: Rad51 replication fork recruitment is required for DNA damage tolerance.
abstract: Homologous recombination (HR) is essential for genome integrity. Recombination proteins participate in tolerating DNA lesions that interfere with DNA replication, but can also generate toxic recombination intermediates and genetic instability when they are not properly regulated. Here, we have studied the role of the recombination proteins Rad51 and Rad52 at replication forks and replicative DNA lesions. We show that Rad52 loads Rad51 onto unperturbed replication forks, where they facilitate replication of alkylated DNA by non-repair functions. The recruitment of Rad52 and Rad51 to chromatin during DNA replication is a prerequisite for the repair of the non-DSB DNA lesions, presumably single-stranded DNA gaps, which are generated during the replication of alkylated DNA. We also show that the repair of these lesions requires CDK1 and is not coupled to the fork but rather restricted to G2/M by the replicative checkpoint. We propose a new scenario for HR where Rad52 and Rad51 are recruited to the fork to promote DNA damage tolerance by distinct and cell cycle-regulated replicative and repair functions.
authors.affiliation: Departamento de Biología Molecular, Centro Andaluz de Biología Molecular y Medicina Regenerativa, Consejo Superior de Investigaciones Científicas, Seville, Spain.
authors.forename: Román
authors.initials: R
authors.lastname: González-Prieto
authors.forename: Ana M
authors.initials: AM
authors.lastname: Muñoz-Cabello
authors.forename: María J
authors.initials: MJ
authors.lastname: Cabello-Lobato
authors.forename: Félix
authors.initials: F
authors.lastname: Prado
mesh_terms: D002843:Chromatin / Q000187:drug effects / Q000378:metabolism; D053903:DNA Breaks, Double-Stranded / Q000187:drug effects; D004249:DNA Damage / Q000187:drug effects / Q000235:genetics; D004260:DNA Repair / Q000235:genetics / Q000502:physiology; D004261:DNA Replication / Q000235:genetics; D004277:DNA, Single-Stranded / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D004706:Endodeoxyribonucleases / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D008836:Micrococcal Nuclease / Q000378:metabolism; D008954:Models, Biological; D011485:Protein Binding / Q000502:physiology; D051135:Rad51 Recombinase / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D051721:Rad52 DNA Repair and Recombination Protein / Q000378:metabolism*; D011995:Recombination, Genetic / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*
chemical_list: D002843:Chromatin; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; C497197:RAD52 protein, S cerevisiae; D051721:Rad52 DNA Repair and Recombination Protein; D029701:Saccharomyces cerevisiae Proteins; C073998:RAD2 protein, S cerevisiae; D008741:Methyl Methanesulfonate; C495151:RAD51 protein, S cerevisiae; D051135:Rad51 Recombinase; D004706:Endodeoxyribonucleases; D008836:Micrococcal Nuclease
references.citation: EMBO J. 2009 Apr 22;28(8):1131-41
references.pmid: 19322196
references.citation: Nat Rev Mol Cell Biol. 2005 Dec;6(12):943-53
references.pmid: 16341080
references.citation: Mol Cell. 2006 Jan 6;21(1):15-27
references.pmid: 16387650
references.citation: Nature. 2002 Sep 12;419(6903):135-41
references.pmid: 12226657
references.citation: EMBO J. 2004 Dec 8;23(24):4868-75
references.pmid: 15549137
references.citation: Annu Rev Genet. 2010;44:113-39
references.pmid: 20690856
references.citation: Nat Struct Mol Biol. 2011 Dec 04;19(1):17-24
references.pmid: 22139015
references.citation: Cell. 2008 Sep 19;134(6):981-94
references.pmid: 18805091
references.citation: PLoS Genet. 2010 Nov 11;6(11):e1001205
references.pmid: 21085632
references.citation: Nature. 2009 Aug 20;460(7258):984-9
references.pmid: 19641493
references.citation: Genes Dev. 2003 Jul 15;17(14):1755-67
references.pmid: 12865299
references.citation: Nat Rev Mol Cell Biol. 2010 Mar;11(3):196-207
references.pmid: 20177395
references.citation: Genetics. 1977 May;86(1):33-55
references.pmid: 195865
references.citation: PLoS Genet. 2006 Nov 10;2(11):e194
references.pmid: 17096599
references.citation: Mol Cell. 2003 May;11(5):1323-36
references.pmid: 12769855
references.citation: EMBO J. 2009 Apr 22;28(8):1121-30
references.pmid: 19262568
references.citation: Annu Rev Biochem. 2008;77:229-57
references.pmid: 18275380
references.citation: Mutat Res. 1977 Apr;43(1):1-10
references.pmid: 194147
references.citation: Mol Cell. 2012 Mar 9;45(5):585-6
references.pmid: 22405272
references.citation: Cell. 2007 Feb 9;128(3):477-90
references.pmid: 17289568
references.citation: Nature. 2005 Jul 21;436(7049):428-33
references.pmid: 15931174
references.citation: J Biol Chem. 1992 Nov 5;267(31):22496-505
references.pmid: 1429600
references.citation: Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8276-82
references.pmid: 11459964
references.citation: PLoS Genet. 2010 May 06;6(5):e1000933
references.pmid: 20463880
references.citation: Cell. 2010 Apr 16;141(2):255-67
references.pmid: 20403322
references.citation: DNA Repair (Amst). 2007 Jul 1;6(7):891-9
references.pmid: 17363342
references.citation: Nature. 2001 Aug 2;412(6846):553-7
references.pmid: 11484057
references.citation: Genes Dev. 2005 Feb 1;19(3):339-50
references.pmid: 15687257
references.citation: Nat Struct Mol Biol. 2011 Aug 14;18(9):1015-9
references.pmid: 21841787
references.citation: Nat Struct Mol Biol. 2010 Nov;17(11):1305-11
references.pmid: 20935632
references.citation: J Cell Biol. 2005 Feb 14;168(4):537-44
references.pmid: 15716375
references.citation: Nature. 2008 Dec 18;456(7224):915-20
references.pmid: 19092928
references.citation: Mol Cell. 2005 Jul 1;19(1):123-33
references.pmid: 15989970
references.citation: Nucleic Acids Res. 2005 Jul 11;33(12):3799-811
references.pmid: 16009812
references.citation: Proc Natl Acad Sci U S A. 2002 Dec 24;99(26):16887-92
references.pmid: 12475932
references.citation: Nat Rev Mol Cell Biol. 2010 Mar;11(3):208-19
references.pmid: 20177396
references.citation: Mol Cell. 2004 Oct 8;16(1):147-57
references.pmid: 15469830
references.citation: J Biol Chem. 1997 Nov 7;272(45):28194-7
references.pmid: 9353267
references.citation: Genes Dev. 2008 Oct 15;22(20):2767-72
references.pmid: 18923075
references.citation: Mol Gen Genet. 1981;184(3):471-8
references.pmid: 7038396
references.citation: Mol Cell. 2005 Sep 2;19(5):699-706
references.pmid: 16137625
references.citation: Cell. 2004 Sep 17;118(6):699-713
references.pmid: 15369670
references.citation: Mol Cell. 2010 Jun 11;38(5):649-61
references.pmid: 20541998
references.citation: Mol Cell Biol. 2007 Nov;27(21):7758-64
references.pmid: 17785441
references.citation: EMBO J. 1995 Aug 1;14(15):3788-99
references.pmid: 7641697
references.citation: Cell. 2011 May 13;145(4):529-42
references.pmid: 21565612
references.citation: Nature. 2010 Jun 17;465(7300):951-5
references.pmid: 20453836
references.citation: Nature. 2004 Oct 21;431(7011):1011-7
references.pmid: 15496928
references.citation: EMBO Rep. 2009 Jul;10(7):790-6
references.pmid: 19465889
references.citation: Nature. 2008 Oct 2;455(7213):689-92
references.pmid: 18716619
references.citation: Nat Cell Biol. 2003 Jun;5(6):572-7
references.pmid: 12766777
references.citation: DNA Repair (Amst). 2008 Oct 1;7(10):1693-704
references.pmid: 18640290
references.citation: Mutat Res. 2011 May-Jun;727(3):104-22
references.pmid: 21558018
references.citation: Nat Cell Biol. 2001 Nov;3(11):958-65
references.pmid: 11715016
references.citation: J Mol Cell Biol. 2011 Feb;3(1):23-30
references.pmid: 21278448
references.citation: Mol Cell. 2010 Feb 26;37(4):492-502
references.pmid: 20188668
references.citation: Cell. 2007 May 18;129(4):665-79
references.pmid: 17512402
references.citation: Nature. 2008 Oct 9;455(7214):770-4
references.pmid: 18806779
references.citation: J Cell Biol. 2009 Jun 15;185(6):969-81
references.pmid: 19506039