Temporal separation of replication and recombination requires the intra-S checkpoint.
In response to DNA damage and replication pausing, eukaryotes activate checkpoint pathways that prevent genomic instability by coordinating cell cycle progression with DNA repair. The intra-S-phase checkpoint has been proposed to protect stalled replication forks from pathological rearrangements that could result from unscheduled recombination. On the other hand, recombination may be needed to cope with either stalled forks or double-strand breaks resulting from hydroxyurea treatment. We have exploited fission yeast to elucidate the relationship between replication fork stalling, loading of replication and recombination proteins onto DNA, and the intra-S checkpoint. Here, we show that a functional recombination machinery is not essential for recovery from replication fork arrest and instead can lead to nonfunctional fork structures. We find that Rad22-containing foci are rare in S-phase cells, but peak in G2 phase cells after a perturbed S phase. Importantly, we find that the intra-S checkpoint is necessary to avoid aberrant strand-exchange events during a hydroxyurea block.
Meister
Peter
P
CNRS UMR2027, Institut Curie, Centre Universitaire, 91405 Orsay Cedex, France.
Taddei
Angela
A
Vernis
Laurence
L
Poidevin
Mickaël
M
Gasser
Susan M
SM
Baldacci
Giuseppe
G
D002470:Cell Survival / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D004249:DNA Damage / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D004260:DNA Repair / Q000235:genetics / Q000502:physiology; D004261:DNA Replication / Q000187:drug effects / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D016195:G2 Phase / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D042822:Genomic Instability / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D006918:Hydroxyurea / Q000633:toxicity; D011995:Recombination, Genetic / Q000187:drug effects / Q000235:genetics*; D016196:S Phase / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D012568:Schizosaccharomyces / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism*
D004268:DNA-Binding Proteins; D029702:Schizosaccharomyces pombe Proteins; C085412:rad52 protein, S pombe; D006918:Hydroxyurea
Nature. 2003 May 15;423(6937):305-9
12748644
Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8276-82
11459964
Genes Cells. 2003 Jul;8(7):573-86
12839619
Mol Cell Biol. 2003 Aug;23(16):5939-46
12897162
Genes Dev. 2000 Jan 1;14(1):81-96
10640278
EMBO J. 2000 Sep 15;19(18):5027-38
10990466
Mol Cell Biol. 2000 Dec;20(23):8758-66
11073977
Nature. 2001 Aug 2;412(6846):553-7
11484057
Nature. 2001 Aug 2;412(6846):557-61
11484058
EMBO J. 2001 Nov 1;20(21):6115-26
11689451
Science. 2002 Jul 26;297(5581):599-602
12142537
J Biol Chem. 2002 Oct 25;277(43):41183-91
12181326
Mol Cell. 2002 Oct;10(4):907-16
12419233
Proc Natl Acad Sci U S A. 2002 Dec 24;99(26):16887-92
12475932
Proc Natl Acad Sci U S A. 2002 Dec 24;99(26):16934-9
12482937
DNA Repair (Amst). 2002 Dec 5;1(12):983-94
12531008
EMBO J. 2003 Aug 15;22(16):4325-36
12912929
Mol Cell Biol. 2003 Sep;23(17):6150-8
12917337
Nucleic Acids Res. 2003 Sep 1;31(17):5064-73
12930957
Nature. 2003 Aug 28;424(6952):1078-83
12944972
Mol Cell Biol. 2003 Nov;23(21):7861-74
14560029
Nat Genet. 2003 Nov;35(3):277-86
14566339
Mutat Res. 2003 Nov 27;532(1-2):59-73
14643429
Genes Dev. 2003 Dec 15;17(24):3017-22
14681210
Mol Cell Biol. 2004 Apr;24(8):3198-212
15060144
Mol Cell Biol. 2004 Apr;24(8):3213-26
15060145
J Cell Biol. 2004 Jun 21;165(6):801-12
15197177
DNA Repair (Amst). 2004 Aug-Sep;3(8-9):835-43
15279768
Chromosoma. 2004 Sep;113(2):92-101
15257466
Mol Biol Cell. 2004 Sep;15(9):4051-63
15229282
Mol Cell Biol. 2004 Oct;24(19):8342-55
15367656
Cell. 2004 Sep 17;118(6):699-713
15369670
Cell. 1987 Nov 6;51(3):463-71
2822257
J Cell Sci. 1990 Nov;97 ( Pt 3):509-16
2074269
Nucleic Acids Res. 1992 Apr 11;20(7):1463-9
1349740
Genes Dev. 1998 Feb 1;12(3):382-95
9450932
Mol Cell Biol. 1999 Jun;19(6):4262-9
10330167
Proc Natl Acad Sci U S A. 1999 Aug 3;96(16):9130-5
10430907
EMBO J. 2005 Jan 26;24(2):405-17
15616582
Nature. 2003 May 15;423(6937):309-12
12748645