Claspin, a Chk1-regulatory protein, monitors DNA replication on chromatin independently of RPA, ATR, and Rad17

被引:166
作者
Lee, J [1 ]
Kumagai, A [1 ]
Dunphy, WG [1 ]
机构
[1] CALTECH, Howard Hughes Med Inst, Div Biol 216 76, Pasadena, CA 91125 USA
关键词
D O I
10.1016/S1097-2765(03)00045-5
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Claspin is required for the ATR-dependent activation of Chk1 in Xenopus egg extracts containing incompletely replicated DNA. We show here that Claspin associates with chromatin in a regulated manner during S phase. Binding of Claspin to chromatin depends on the pre-replication complex (pre-RC) and Cdc45 but not on replication protein A (RPA). These dependencies suggest that binding of Claspin occurs around the time of initial DNA unwinding at replication origins. By contrast, both ATR and Rad17 require RPA for association with DNA. Claspin, ATR, and Rad17 all bind to chromatin independently. These findings suggest that Claspin plays a role in monitoring DNA replication during S phase. Claspin, ATR, and Rad17 may collaborate in checkpoint regulation by detecting different aspects of a DNA replication fork.
引用
收藏
页码:329 / 340
页数:12
相关论文
共 39 条
[1]   Cell cycle checkpoint signaling through the ATM and ATR kinases [J].
Abraham, RT .
GENES & DEVELOPMENT, 2001, 15 (17) :2177-2196
[2]   Mrc1 transduces signals of DNA replication stress to activate Rad53 [J].
Alcasabas, AA ;
Osborn, AJ ;
Bachant, J ;
Hu, FH ;
Werler, PJH ;
Bousset, K ;
Furuya, K ;
Diffley, JFX ;
Carr, AM ;
Elledge, SJ .
NATURE CELL BIOLOGY, 2001, 3 (11) :958-965
[3]   ATR/ATM-mediated phosphorylation of human Rad17 is required for genotoxic stress responses [J].
Bao, SD ;
Tibbetts, RS ;
Brumbaugh, KM ;
Fang, YN ;
Richardson, DA ;
Ali, A ;
Chen, SM ;
Abraham, RT ;
Wang, XF .
NATURE, 2001, 411 (6840) :969-974
[4]   Chk2 kinase - A busy messenger [J].
Bartek, J ;
Falck, J ;
Lukas, J .
NATURE REVIEWS MOLECULAR CELL BIOLOGY, 2001, 2 (12) :877-886
[5]   DNA replication in eukaryotic cells [J].
Bell, SP ;
Dutta, A .
ANNUAL REVIEW OF BIOCHEMISTRY, 2002, 71 :333-374
[6]   Characterization of Schizosaccharomyces pombe Hus1:: a PCNA-related protein that associates with Rad1 and Rad9 [J].
Caspari, T ;
Dahlen, M ;
Kanter-Smoler, G ;
Lindsay, HD ;
Hofmann, K ;
Papadimitriou, K ;
Sunnerhagen, P ;
Carr, AM .
MOLECULAR AND CELLULAR BIOLOGY, 2000, 20 (04) :1254-1262
[7]   SEPARATE DOMAINS OF P21 INVOLVED IN THE INHIBITION OF CDK KINASE AND PCNA [J].
CHEN, JJ ;
JACKSON, PK ;
KIRSCHNER, MW ;
DUTTA, A .
NATURE, 1995, 374 (6520) :386-388
[8]   The Xenopus Cdc6 protein is essential for the initiation of a single round of DNA replication in cell-free extracts [J].
Coleman, TR ;
Carpenter, PB ;
Dunphy, WG .
CELL, 1996, 87 (01) :53-63
[9]   FISSION YEAST RAD17 - A HOMOLOG OF BUDDING YEAST RAD24 THAT SHARES REGIONS OF SEQUENCE SIMILARITY WITH DNA-POLYMERASE ACCESSORY PROTEINS [J].
GRIFFITHS, DJF ;
BARBET, NC ;
MCCREADY, S ;
LEHMANN, AR ;
CARR, AM .
EMBO JOURNAL, 1995, 14 (23) :5812-5823
[10]   Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts [J].
Guo, ZJ ;
Kumagai, A ;
Wang, SX ;
Dunphy, WG .
GENES & DEVELOPMENT, 2000, 14 (21) :2745-2756