Abstract
Using the Xenopus egg extract system, we investigated the involvement of DNA replication in activation of the DNA damage checkpoint. We show here that DNA damage slows replication in a checkpoint-independent manner and is accompanied by replication-dependent recruitment of ATR and Rad1 to chromatin. We also find that the replication proteins RPA and Polalpha accumulate on chromatin following DNA damage. Finally, damage-induced Chk1 phosphorylation and checkpoint arrest are abrogated when replication is inhibited. These data indicate that replication is required for activation of the DNA damage checkpoint and suggest a unifying model for ATR activation by diverse lesions during S phase.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Ataxia Telangiectasia Mutated Proteins
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Cell Cycle
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Cell Cycle Proteins / metabolism*
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Checkpoint Kinase 1
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DNA Damage*
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DNA Replication* / radiation effects
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DNA-Binding Proteins*
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Endonucleases / genetics
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Endonucleases / metabolism
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Female
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In Vitro Techniques
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Methyl Methanesulfonate / toxicity
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Models, Biological
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Oocytes / drug effects
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Oocytes / metabolism
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Oocytes / radiation effects
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Protein Kinases / metabolism
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Protein Serine-Threonine Kinases*
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Ultraviolet Rays / adverse effects
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Xenopus
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Xenopus Proteins*
Substances
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Cell Cycle Proteins
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DNA-Binding Proteins
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Recombinant Proteins
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Xenopus Proteins
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Methyl Methanesulfonate
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Protein Kinases
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Atr protein, Xenopus
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Ataxia Telangiectasia Mutated Proteins
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Checkpoint Kinase 1
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Chek1 protein, Xenopus
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Protein Serine-Threonine Kinases
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Endonucleases