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.
Phosphorylation of Xenopus Rad1 and Hus1 defines a readout for ATR activation that is independent of Claspin and the Rad9 carboxy terminus.
Lupardus PJ, Cimprich KA.
???displayArticle.abstract???
The DNA damage checkpoint pathways sense and respond to DNA damage to ensure genomic stability. The ATR kinase is a central regulator of one such pathway and phosphorylates a number of proteins that have roles in cell cycle progression and DNA repair. Using the Xenopus egg extract system, we have investigated regulation of the Rad1/Hus1/Rad9 complex. We show here that phosphorylation of Rad1 and Hus1 occurs in an ATR- and TopBP1-dependent manner on T5 of Rad1 and S219 and T223 of Hus1. Mutation of these sites has no effect on the phosphorylation of Chk1 by ATR. Interestingly, phosphorylation of Rad1 is independent of Claspin and the Rad9 carboxy terminus, both of which are required for Chk1 phosphorylation. These data suggest that an active ATR signaling complex exists in the absence of the carboxy terminus of Rad9 and that this carboxy-terminal domain may be a specific requirement for Chk1 phosphorylation and not necessary for all ATR-mediated signaling events. Thus, Rad1 phosphorylation provides an alternate and early readout for the study of ATR activation.
Bell,
DNA replication in eukaryotic cells.
2002, Pubmed
Bell,
DNA replication in eukaryotic cells.
2002,
Pubmed Bermudez,
Loading of the human 9-1-1 checkpoint complex onto DNA by the checkpoint clamp loader hRad17-replication factor C complex in vitro.
2003,
Pubmed Brown,
ATR disruption leads to chromosomal fragmentation and early embryonic lethality.
2000,
Pubmed Burtelow,
Retention of the human Rad9 checkpoint complex in extraction-resistant nuclear complexes after DNA damage.
2000,
Pubmed Burtelow,
Reconstitution and molecular analysis of the hRad9-hHus1-hRad1 (9-1-1) DNA damage responsive checkpoint complex.
2001,
Pubmed Byun,
Functional uncoupling of MCM helicase and DNA polymerase activities activates the ATR-dependent checkpoint.
2005,
Pubmed
,
Xenbase Chang,
Interaction of checkpoint proteins Hus1/Rad1/Rad9 with DNA base excision repair enzyme MutY homolog in fission yeast, Schizosaccharomyces pombe.
2005,
Pubmed Chen,
Chk1 in the DNA damage response: conserved roles from yeasts to mammals.
2004,
Pubmed Cimprich,
Fragile sites: breaking up over a slowdown.
2003,
Pubmed Cortez,
Unwind and slow down: checkpoint activation by helicase and polymerase uncoupling.
2005,
Pubmed Cortez,
ATR and ATRIP: partners in checkpoint signaling.
2001,
Pubmed Costanzo,
An ATR- and Cdc7-dependent DNA damage checkpoint that inhibits initiation of DNA replication.
2003,
Pubmed
,
Xenbase Edwards,
A Rad3-Rad26 complex responds to DNA damage independently of other checkpoint proteins.
1999,
Pubmed Ellison,
Biochemical characterization of DNA damage checkpoint complexes: clamp loader and clamp complexes with specificity for 5' recessed DNA.
2003,
Pubmed Furstenthal,
Cyclin E uses Cdc6 as a chromatin-associated receptor required for DNA replication.
2001,
Pubmed
,
Xenbase Furuya,
Chk1 activation requires Rad9 S/TQ-site phosphorylation to promote association with C-terminal BRCT domains of Rad4TOPBP1.
2004,
Pubmed Garcia,
Identification and functional analysis of TopBP1 and its homologs.
2005,
Pubmed Greer,
hRad9 rapidly binds DNA containing double-strand breaks and is required for damage-dependent topoisomerase II beta binding protein 1 focus formation.
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 Hopkins,
Deletion of mouse rad9 causes abnormal cellular responses to DNA damage, genomic instability, and embryonic lethality.
2004,
Pubmed Jones,
XRad17 is required for the activation of XChk1 but not XCds1 during checkpoint signaling in Xenopus.
2003,
Pubmed
,
Xenbase Kai,
Checkpoint activation regulates mutagenic translesion synthesis.
2003,
Pubmed Kelly,
Regulation of chromosome replication.
2000,
Pubmed Kim,
Substrate specificities and identification of putative substrates of ATM kinase family members.
1999,
Pubmed Kostrub,
Hus1p, a conserved fission yeast checkpoint protein, interacts with Rad1p and is phosphorylated in response to DNA damage.
1998,
Pubmed 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 Kumagai,
Claspin and the activated form of ATR-ATRIP collaborate in the activation of Chk1.
2004,
Pubmed
,
Xenbase Lee,
Claspin, a Chk1-regulatory protein, monitors DNA replication on chromatin independently of RPA, ATR, and Rad17.
2003,
Pubmed
,
Xenbase Li,
Absence of BLM leads to accumulation of chromosomal DNA breaks during both unperturbed and disrupted S phases.
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 Lupardus,
A requirement for replication in activation of the ATR-dependent DNA damage checkpoint.
2002,
Pubmed
,
Xenbase Mäkiniemi,
BRCT domain-containing protein TopBP1 functions in DNA replication and damage response.
2001,
Pubmed Melo,
A unified view of the DNA-damage checkpoint.
2002,
Pubmed
,
Xenbase Murray,
Cell cycle extracts.
1991,
Pubmed Parrilla-Castellar,
Dial 9-1-1 for DNA damage: the Rad9-Hus1-Rad1 (9-1-1) clamp complex.
2004,
Pubmed Parrilla-Castellar,
Cut5 is required for the binding of Atr and DNA polymerase alpha to genotoxin-damaged chromatin.
2003,
Pubmed
,
Xenbase Peng,
Mitotic and G2 checkpoint control: regulation of 14-3-3 protein binding by phosphorylation of Cdc25C on serine-216.
1997,
Pubmed Roos-Mattjus,
Phosphorylation of human Rad9 is required for genotoxin-activated checkpoint signaling.
2003,
Pubmed Sanchez,
Conservation of the Chk1 checkpoint pathway in mammals: linkage of DNA damage to Cdk regulation through Cdc25.
1997,
Pubmed Sarkaria,
Inhibition of ATM and ATR kinase activities by the radiosensitizing agent, caffeine.
1999,
Pubmed Smirnova,
The human checkpoint sensor and alternative DNA clamp Rad9-Rad1-Hus1 modulates the activity of DNA ligase I, a component of the long-patch base excision repair machinery.
2005,
Pubmed Stokes,
DNA replication is required for the checkpoint response to damaged DNA in Xenopus egg extracts.
2002,
Pubmed
,
Xenbase St Onge,
A role for the phosphorylation of hRad9 in checkpoint signaling.
2003,
Pubmed Toueille,
The human Rad9/Rad1/Hus1 damage sensor clamp interacts with DNA polymerase beta and increases its DNA substrate utilisation efficiency: implications for DNA repair.
2004,
Pubmed Venclovas,
Structure-based predictions of Rad1, Rad9, Hus1 and Rad17 participation in sliding clamp and clamp-loading complexes.
2000,
Pubmed Wang,
Genetic and physical interactions between DPB11 and DDC1 in the yeast DNA damage response pathway.
2002,
Pubmed Wang,
The human Rad9-Rad1-Hus1 checkpoint complex stimulates flap endonuclease 1.
2004,
Pubmed Weiss,
Inactivation of mouse Hus1 results in genomic instability and impaired responses to genotoxic stress.
2000,
Pubmed Weiss,
Critical role for mouse Hus1 in an S-phase DNA damage cell cycle checkpoint.
2003,
Pubmed 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,
ATR-mediated checkpoint pathways regulate phosphorylation and activation of human Chk1.
2001,
Pubmed Zhou,
The DNA damage response: putting checkpoints in perspective.
2000,
Pubmed Zou,
Sensing DNA damage through ATRIP recognition of RPA-ssDNA complexes.
2003,
Pubmed Zou,
Replication protein A-mediated recruitment and activation of Rad17 complexes.
2003,
Pubmed Zou,
Regulation of ATR substrate selection by Rad17-dependent loading of Rad9 complexes onto chromatin.
2002,
Pubmed
