Lupardus Patrick J, Byun Tony, Yee Muh-Ching, Hekmat-Nejad Mohammad, Cimprich Karlene A
Department of Molecular Pharmacology, Stanford University, Stanford, California 94305-5174, USA.
Genes Dev. 2002 Sep 15;16(18):2327-32. doi: 10.1101/gad.1013502.
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.
利用非洲爪蟾卵提取物系统,我们研究了DNA复制在DNA损伤检查点激活过程中的作用。我们在此表明,DNA损伤以一种不依赖检查点的方式减缓复制,并伴随着ATR和Rad1依赖复制的向染色质募集。我们还发现,DNA损伤后,复制蛋白RPA和Polα在染色质上积累。最后,当复制受到抑制时,损伤诱导的Chk1磷酸化和检查点停滞被消除。这些数据表明,DNA损伤检查点的激活需要复制,并提示了一个关于S期不同损伤激活ATR的统一模型。