Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Mol Biol Cell
2005 Nov 01;1611:5269-82. doi: 10.1091/mbc.e05-07-0671.
Show Gene links
Show Anatomy links
Roles of replication fork-interacting and Chk1-activating domains from Claspin in a DNA replication checkpoint response.
Lee J
,
Gold DA
,
Shevchenko A
,
Shevchenko A
,
Dunphy WG
.
???displayArticle.abstract??? Claspin is essential for the ATR-dependent activation of Chk1 in Xenopus egg extracts containing incompletely replicated DNA. Claspin associates with replication forks upon origin unwinding. We show that Claspin contains a replication fork-interacting domain (RFID, residues 265-605) that associates with Cdc45, DNA polymerase epsilon, replication protein A, and two replication factor C complexes on chromatin. The RFID contains two basic patches (BP1 and BP2) at amino acids 265-331 and 470-600, respectively. Deletion of either BP1 or BP2 compromises optimal binding of Claspin to chromatin. Absence of BP1 has no effect on the ability of Claspin to mediate activation of Chk1. By contrast, removal of BP2 causes a large reduction in the Chk1-activating potency of Claspin. We also find that Claspin contains a small Chk1-activating domain (residues 776-905) that does not bind stably to chromatin, but it is fully effective at high concentrations for mediating activation of Chk1. These results indicate that stable retention of Claspin on chromatin is not necessary for activation of Chk1. Instead, our findings suggest that only transient interaction of Claspin with replication forks potentiates its Chk1-activating function. Another implication of this work is that stable binding of Claspin to chromatin may play a role in other functions besides the activation of Chk1.
Abraham,
Cell cycle checkpoint signaling through the ATM and ATR kinases.
2001, Pubmed
Abraham,
Cell cycle checkpoint signaling through the ATM and ATR kinases.
2001,
Pubmed
Alcasabas,
Mrc1 transduces signals of DNA replication stress to activate Rad53.
2001,
Pubmed
Canman,
Checkpoint mediators: relaying signals from DNA strand breaks.
2003,
Pubmed
Celeste,
Histone H2AX phosphorylation is dispensable for the initial recognition of DNA breaks.
2003,
Pubmed
Chini,
Human claspin is required for replication checkpoint control.
2003,
Pubmed
,
Xenbase
Cortez,
ATR and ATRIP: partners in checkpoint signaling.
2001,
Pubmed
Ellison,
Biochemical characterization of DNA damage checkpoint complexes: clamp loader and clamp complexes with specificity for 5' recessed DNA.
2003,
Pubmed
Guo,
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.
2000,
Pubmed
,
Xenbase
Hekmat-Nejad,
Xenopus ATR is a replication-dependent chromatin-binding protein required for the DNA replication checkpoint.
,
Pubmed
,
Xenbase
Huyen,
Methylated lysine 79 of histone H3 targets 53BP1 to DNA double-strand breaks.
2004,
Pubmed
Jeong,
Phosphorylated claspin interacts with a phosphate-binding site in the kinase domain of Chk1 during ATR-mediated activation.
2003,
Pubmed
,
Xenbase
Katou,
S-phase checkpoint proteins Tof1 and Mrc1 form a stable replication-pausing complex.
2003,
Pubmed
Kumagai,
Claspin and the activated form of ATR-ATRIP collaborate in the activation of Chk1.
2004,
Pubmed
,
Xenbase
Kumagai,
Claspin, a novel protein required for the activation of Chk1 during a DNA replication checkpoint response in Xenopus egg extracts.
2000,
Pubmed
,
Xenbase
Kumagai,
The Xenopus Chk1 protein kinase mediates a caffeine-sensitive pathway of checkpoint control in cell-free extracts.
1998,
Pubmed
,
Xenbase
Kumagai,
Repeated phosphopeptide motifs in Claspin mediate the regulated binding of Chk1.
2003,
Pubmed
,
Xenbase
Lee,
Claspin, a Chk1-regulatory protein, monitors DNA replication on chromatin independently of RPA, ATR, and Rad17.
2003,
Pubmed
,
Xenbase
Lin,
Human Claspin works with BRCA1 to both positively and negatively regulate cell proliferation.
2004,
Pubmed
,
Xenbase
Liu,
Chk1 is an essential kinase that is regulated by Atr and required for the G(2)/M DNA damage checkpoint.
2000,
Pubmed
Marheineke,
Control of replication origin density and firing time in Xenopus egg extracts: role of a caffeine-sensitive, ATR-dependent checkpoint.
2004,
Pubmed
,
Xenbase
McGowan,
The DNA damage response: sensing and signaling.
2004,
Pubmed
Melo,
A unified view of the DNA-damage checkpoint.
2002,
Pubmed
,
Xenbase
Michael,
Activation of the DNA replication checkpoint through RNA synthesis by primase.
2000,
Pubmed
,
Xenbase
Mimura,
Xenopus Cdc45-dependent loading of DNA polymerase alpha onto chromatin under the control of S-phase Cdk.
1998,
Pubmed
,
Xenbase
Mimura,
Central role for cdc45 in establishing an initiation complex of DNA replication in Xenopus egg extracts.
2000,
Pubmed
,
Xenbase
Moore,
Identification of the nuclear localization signal in Xenopus cyclin E and analysis of its role in replication and mitosis.
2002,
Pubmed
,
Xenbase
Nakamura,
Histone H2A phosphorylation controls Crb2 recruitment at DNA breaks, maintains checkpoint arrest, and influences DNA repair in fission yeast.
2004,
Pubmed
O'Connell,
G2 damage checkpoints: what is the turn-on?
2005,
Pubmed
Osborn,
Mrc1 is a replication fork component whose phosphorylation in response to DNA replication stress activates Rad53.
2003,
Pubmed
Osborn,
Checking on the fork: the DNA-replication stress-response pathway.
2002,
Pubmed
Sancar,
Molecular mechanisms of mammalian DNA repair and the DNA damage checkpoints.
2004,
Pubmed
Sanders,
Methylation of histone H4 lysine 20 controls recruitment of Crb2 to sites of DNA damage.
2004,
Pubmed
Sar,
Human claspin is a ring-shaped DNA-binding protein with high affinity to branched DNA structures.
2004,
Pubmed
,
Xenbase
Shechter,
ATR and ATM regulate the timing of DNA replication origin firing.
2004,
Pubmed
,
Xenbase
Shevchenko,
Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels.
1996,
Pubmed
Shevchenko,
Charting the proteomes of organisms with unsequenced genomes by MALDI-quadrupole time-of-flight mass spectrometry and BLAST homology searching.
2001,
Pubmed
Shevchenko,
Nanoelectrospray tandem mass spectrometry and sequence similarity searching for identification of proteins from organisms with unknown genomes.
2003,
Pubmed
Waga,
DNA polymerase epsilon is required for coordinated and efficient chromosomal DNA replication in Xenopus egg extracts.
2001,
Pubmed
,
Xenbase
Xu,
Mrc1 is required for sister chromatid cohesion to aid in recombination repair of spontaneous damage.
2004,
Pubmed
Yanow,
Xenopus Drf1, a regulator of Cdc7, displays checkpoint-dependent accumulation on chromatin during an S-phase arrest.
2003,
Pubmed
,
Xenbase
Yoo,
Adaptation of a DNA replication checkpoint response depends upon inactivation of Claspin by the Polo-like kinase.
2004,
Pubmed
,
Xenbase
You,
The role of single-stranded DNA and polymerase alpha in establishing the ATR, Hus1 DNA replication checkpoint.
2002,
Pubmed
,
Xenbase
Zhao,
DNA binding domain in the replication checkpoint protein Mrc1 of Schizosaccharomyces pombe.
2004,
Pubmed
Zou,
Replication protein A-mediated recruitment and activation of Rad17 complexes.
2003,
Pubmed
