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Cell Cycle
2011 Feb 01;103:500-6. doi: 10.4161/cc.10.3.14753.
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A new in vitro system for activating the cell cycle checkpoint.
Wang J
,
Engle S
,
Zhang Y
.
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In response to DNA damage, cells launch elegant networks of genome surveillance mechanisms, called cell cycle checkpoints, to detect and repair damaged DNA to maintain the genome stability. Key components of cell cycle checkpoints are two PI3K-related protein kinases (PIKK), ATR and ATM, which participate in both sensing the DNA damage and transducing the damage signal through phosphorylating two target protein kinases, Chk1 and Chk2, respectively. However, how exactly cell cycle checkpoints are activated, maintained, and terminated are not completely understood. Given the complexity of the cell cycle checkpoint signaling and the cellular environment, systems that can faithfully mimic the cell cycle checkpoint activation in vitro, such as the Xenopus egg extracts, are of extreme value in dissecting the precise molecular mechanisms underlying DNA damage response. Here we describe that the well-established in vitro transcription and translation (IVTNT) system has the capability to induce protein phosphorylation of substrates for ATR or ATM, including Chk1, Rad17, and ATM itself. These phosphorylation events highly mimic those occurring in cells when treated with DNA damaging agents. Our results demonstrate that the IVTNT system could be developed into a novel in vitro system to facilitating the dissecting of mechanisms leading to cell cycle checkpoint activation in vivo.
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
Bakkenist,
DNA damage activates ATM through intermolecular autophosphorylation and dimer dissociation.
2003,
Pubmed
Bao,
ATR/ATM-mediated phosphorylation of human Rad17 is required for genotoxic stress responses.
2001,
Pubmed
Bartek,
DNA damage checkpoints: from initiation to recovery or adaptation.
2007,
Pubmed
Bartkova,
DNA damage response as a candidate anti-cancer barrier in early human tumorigenesis.
2005,
Pubmed
Blasina,
Breaching the DNA damage checkpoint via PF-00477736, a novel small-molecule inhibitor of checkpoint kinase 1.
2008,
Pubmed
Brumbaugh,
The mRNA surveillance protein hSMG-1 functions in genotoxic stress response pathways in mammalian cells.
2004,
Pubmed
Byun,
Functional uncoupling of MCM helicase and DNA polymerase activities activates the ATR-dependent checkpoint.
2005,
Pubmed
,
Xenbase
Casper,
ATR regulates fragile site stability.
2002,
Pubmed
Cha,
ATR homolog Mec1 promotes fork progression, thus averting breaks in replication slow zones.
2002,
Pubmed
Chini,
Human claspin is required for replication checkpoint control.
2003,
Pubmed
,
Xenbase
Choi,
Cooperative activation of the ATR checkpoint kinase by TopBP1 and damaged DNA.
2009,
Pubmed
Choi,
The human ATR-mediated DNA damage checkpoint in a reconstituted system.
2009,
Pubmed
Choi,
Reconstitution of RPA-covered single-stranded DNA-activated ATR-Chk1 signaling.
2010,
Pubmed
Clarke,
DNA-dependent phosphorylation of Chk1 and Claspin in a human cell-free system.
2005,
Pubmed
,
Xenbase
Cortez,
ATR and ATRIP: partners in checkpoint signaling.
2001,
Pubmed
Cuadrado,
ATM regulates ATR chromatin loading in response to DNA double-strand breaks.
2006,
Pubmed
Delacroix,
The Rad9-Hus1-Rad1 (9-1-1) clamp activates checkpoint signaling via TopBP1.
2007,
Pubmed
Garner,
Studying the DNA damage response using in vitro model systems.
2009,
Pubmed
,
Xenbase
Gorgoulis,
Activation of the DNA damage checkpoint and genomic instability in human precancerous lesions.
2005,
Pubmed
Harper,
The DNA damage response: ten years after.
2007,
Pubmed
Jazayeri,
ATM- and cell cycle-dependent regulation of ATR in response to DNA double-strand breaks.
2006,
Pubmed
Kastan,
Cell-cycle checkpoints and cancer.
2004,
Pubmed
Katzen,
The past, present and future of cell-free protein synthesis.
2005,
Pubmed
Kim,
Phosphorylation of Chk1 by ATM- and Rad3-related (ATR) in Xenopus egg extracts requires binding of ATRIP to ATR but not the stable DNA-binding or coiled-coil domains of ATRIP.
2005,
Pubmed
,
Xenbase
Kumagai,
TopBP1 activates the ATR-ATRIP complex.
2006,
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
Lee,
Direct activation of the ATM protein kinase by the Mre11/Rad50/Nbs1 complex.
2004,
Pubmed
Li,
Regulation of CHK2 by DNA-dependent protein kinase.
2005,
Pubmed
Liu,
Chk1 is an essential kinase that is regulated by Atr and required for the G(2)/M DNA damage checkpoint.
2000,
Pubmed
Lovejoy,
Common mechanisms of PIKK regulation.
2009,
Pubmed
Myers,
Rapid activation of ATR by ionizing radiation requires ATM and Mre11.
2006,
Pubmed
Niida,
Specific role of Chk1 phosphorylations in cell survival and checkpoint activation.
2007,
Pubmed
Osborn,
Checking on the fork: the DNA-replication stress-response pathway.
2002,
Pubmed
Pacek,
A requirement for MCM7 and Cdc45 in chromosome unwinding during eukaryotic DNA replication.
2004,
Pubmed
,
Xenbase
Sarkaria,
Inhibition of ATM and ATR kinase activities by the radiosensitizing agent, caffeine.
1999,
Pubmed
Sartori,
Human CtIP promotes DNA end resection.
2007,
Pubmed
Shiloh,
The ATM-mediated DNA-damage response: taking shape.
2006,
Pubmed
,
Xenbase
Shiotani,
Single-stranded DNA orchestrates an ATM-to-ATR switch at DNA breaks.
2009,
Pubmed
Takeda,
Ctp1/CtIP and the MRN complex collaborate in the initial steps of homologous recombination.
2007,
Pubmed
Unsal-Kaçmaz,
The human Tim/Tipin complex coordinates an Intra-S checkpoint response to UV that slows replication fork displacement.
2007,
Pubmed
Walker,
Chk1 C-terminal regulatory phosphorylation mediates checkpoint activation by de-repression of Chk1 catalytic activity.
2009,
Pubmed
Wang,
Rad17 phosphorylation is required for claspin recruitment and Chk1 activation in response to replication stress.
2006,
Pubmed
Wilsker,
Essential function of Chk1 can be uncoupled from DNA damage checkpoint and replication control.
2008,
Pubmed
Xu,
Chk2 activation and phosphorylation-dependent oligomerization.
2002,
Pubmed
You,
Rapid activation of ATM on DNA flanking double-strand breaks.
2007,
Pubmed
,
Xenbase
Zhang,
Genotoxic stress targets human Chk1 for degradation by the ubiquitin-proteasome pathway.
2005,
Pubmed
Zhang,
The F box protein Fbx6 regulates Chk1 stability and cellular sensitivity to replication stress.
2009,
Pubmed
Zhao,
ATR-mediated checkpoint pathways regulate phosphorylation and activation of human Chk1.
2001,
Pubmed
Zou,
Sensing DNA damage through ATRIP recognition of RPA-ssDNA complexes.
2003,
Pubmed
Zou,
Regulation of ATR substrate selection by Rad17-dependent loading of Rad9 complexes onto chromatin.
2002,
Pubmed
