Abstract
Cells maintain genomic stability by the coordination of DNA-damage repair and cell-cycle checkpoint control. In replicating cells, DNA damage usually activates intra-S-phase checkpoint controls, which are characterized by delayed S-phase progression and increased Rad53 phosphorylation. We show that in budding yeast, the intra-S-phase checkpoint controls, although functional, are not activated by the topoisomerase I inhibitor camptothecin (CPT). In a CPT-hypersensitive mutant strain that lacks the histone 2A (H2A) phosphatidylinositol-3-OH kinase (PI(3)K) motif at Ser 129 (h2a-s129a), the hypersensitivity was found to result from a failure to process full-length chromosomal DNA molecules during ongoing replication. H2A Ser 129 is not epistatic to the RAD24 and RAD9 checkpoint genes, suggesting a non-checkpoint role for the H2A PI(3)K site. These results suggest that H2A Ser 129 is an essential component for the efficient repair of DNA double-stranded breaks (DSBs) during replication in yeast, particularly of those DSBs that do not induce the intra-S-phase checkpoint.
MeSH Terms
Animals
Camptothecin/pharmacology
Cell Cycle/physiology
Chromosomes, Fungal/genetics,metabolism,radiation effects
DNA Damage/genetics,radiation effects
DNA Repair/physiology
DNA Topoisomerases, Type I/metabolism
Electrophoresis, Gel, Pulsed-Field
Histones/chemistry,genetics,metabolism
Humans
Mutation, Missense/genetics
Saccharomyces cerevisiae/cytology,genetics,metabolism,radiation effects
Serine/genetics,metabolism
Topoisomerase I Inhibitors
Chemicals
Histones
Topoisomerase I Inhibitors
Serine
DNA Topoisomerases, Type I
Camptothecin
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Redon Christophe
Laboratory of Molecular Pharmacology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
Pilch Duane R
Rogakou Emmy P
Orr Ann H
Lowndes Noel F
Bonner William M
References (15)
15 references, click to expand
-
Topoisomerase I-mediated DNA damage.
Adv Cancer Res. 2001;80:189-216
PMID: 11034544
-
A role for Saccharomyces cerevisiae histone H2A in DNA repair.
Nature. 2000 Dec 21-28;408(6815):1001-4
PMID: 11140636
-
A critical role for histone H2AX in recruitment of repair factors to nuclear foci after DNA damage.
Curr Biol. 2000 Jul 27-Aug 10;10(15):886-95
PMID: 10959836
-
Megabase chromatin domains involved in DNA double-strand breaks in vivo.
J Cell Biol. 1999 Sep 6;146(5):905-16
PMID: 10477747
-
Regulation of DNA-replication origins during cell-cycle progression.
Nature. 1998 Oct 8;395(6702):618-21
PMID: 9783590
-
Recovery from DNA replicational stress is the essential function of the S-phase checkpoint pathway.
Genes Dev. 1998 Sep 15;12(18):2956-70
PMID: 9744871
-
DNA damage checkpoints update: getting molecular.
Curr Opin Genet Dev. 1998 Apr;8(2):185-93
PMID: 9610409
-
RAD9 and RAD24 define two additive, interacting branches of the DNA damage checkpoint pathway in budding yeast normally required for Rad53 modification and activation.
EMBO J. 1998 May 1;17(9):2687-98
PMID: 9564050
-
DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139.
J Biol Chem. 1998 Mar 6;273(10):5858-68
PMID: 9488723
-
The yeast CDC16 and CDC27 genes restrict DNA replication to once per cell cycle.
Cell. 1996 Apr 5;85(1):39-48
PMID: 8620535
-
Regulation of RAD53 by the ATM-like kinases MEC1 and TEL1 in yeast cell cycle checkpoint pathways.
Science. 1996 Jan 19;271(5247):357-60
PMID: 8553072
-
Structure-activity study of the actions of camptothecin derivatives on mammalian topoisomerase I: evidence for a specific receptor site and a relation to antitumor activity.
Cancer Res. 1989 Mar 15;49(6):1465-9
PMID: 2538227
-
ATM and ATR: networking cellular responses to DNA damage.
Curr Opin Genet Dev. 2001 Feb;11(1):71-7
PMID: 11163154
-
Conversion of topoisomerase I cleavage complexes on the leading strand of ribosomal DNA into 5'-phosphorylated DNA double-strand breaks by replication runoff.
Mol Cell Biol. 2000 Jun;20(11):3977-87
PMID: 10805740
-
Genomic instability in mice lacking histone H2AX.
Science. 2002 May 3;296(5569):922-7
PMID: 11934988