Human DDK rescues stalled forks and counteracts checkpoint inhibition at unfired origins to complete DNA replication.
Human DDK rescues stalled forks and counteracts checkpoint inhibition at unfired origins to complete DNA replication.
复制标题
人类DDK拯救停滞的分叉和抵消检查点抑制在未启动的起源完成DNA复制。
DOI：
10.1016/j.molcel.2021.01.004
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发表时间：
2021-02-04
期刊：
影响因子：
16
通讯作者：
Jallepalli PV
中科院分区：
文献类型：
--
作者：
Jones MJK;Gelot C;Munk S;Koren A;Kawasoe Y;George KA;Santos RE;Olsen JV;McCarroll SA;Frattini MG;Takahashi TS;Jallepalli PV
真核生物基因组复制通过空间和时间调控的起源发射。细胞周期蛋白依赖性激酶（CDK）和Dbf 4依赖性激酶（DDK）促进起源放电，而S期检查点限制放电以防止核苷酸和RPA耗尽。我们使用化学遗传学以最大精度询问人类DDK，剖析其与S期检查点的关系，并确定DDK底物。我们表明，DDK抑制（DDKi）会导致起源放电和分叉停滞的分级抑制。S期检查点抑制拯救了DDKi细胞和DDK耗尽的爪蟾卵提取物中的起源放电。DDK抑制还损害RPA加载、新生链保护和分叉重启。通过定量磷酸蛋白质组学，我们确定BRCA 1-A复合物亚基MERIT 40和粘附素辅助亚基PDS 5 B作为DDK效应子在叉保护和重启。磷酸化中和两种底物中由固有无序区域介导的自抑制。我们的研究结果揭示了DDK控制大型脊椎动物基因组复制的机制。真核生物基因组通过复制起点的空间和时间调控的发射进行复制。利用化学遗传学和磷酸蛋白质组学，Jones等人表明，人类DDK通过抵消S期检查点来促进起源放电，并且还能够解偶联，保护和重新启动停滞的分叉。
Eukaryotic genomes replicate via spatially and temporally regulated origin firing. Cyclin-dependent kinase (CDK) and Dbf4-dependent kinase (DDK) promote origin firing, whereas the S phase checkpoint limits firing to prevent nucleotide and RPA exhaustion. We used chemical genetics to interrogate human DDK with maximum precision, dissect its relationship with the S phase checkpoint, and identify DDK substrates. We show that DDK inhibition (DDKi) leads to graded suppression of origin firing and fork arrest. S phase checkpoint inhibition rescued origin firing in DDKi cells and DDK-depleted Xenopus egg extracts. DDK inhibition also impairs RPA loading, nascent strand protection, and fork restart. Via quantitative phosphoproteomics, we identify the BRCA1-A complex subunit MERIT40 and the cohesin accessory subunit PDS5B as DDK effectors in fork protection and restart. Phosphorylation neutralizes auto-inhibition mediated by intrinsically disordered regions in both substrates. Our results reveal mechanisms through which DDK controls the duplication of large vertebrate genomes. Eukaryote genomes duplicate via spatially and temporally regulated firing of replication origins. Using chemical genetics and phosphoproteomics, Jones et al. show that human DDK promotes origin firing by counteracting the S phase checkpoint, and also enables the uncoupling, protection, and restart of stalled forks.
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影响因子：
7.5
作者：
Fu H;Baris A;Aladjem MI
通讯作者：
Aladjem MI
影响因子：
16.8
作者：
Bhat KP;Cortez D
通讯作者：
Cortez D
DOI：
10.1073/pnas.0911500106
发表时间：
2009-12-01
影响因子：
11.1
作者：
Evrin, Cecile;Clarke, Pippa;Speck, Christian
通讯作者：
Speck, Christian
影响因子：
3.3
作者：
Chen, Ying-Chou;Kenworthy, Jessica;Weinreich, Michael
通讯作者：
Weinreich, Michael
影响因子：
8.8
作者：
Bainor, Anthony J.;Saini, Siddharth;David, Gregory
通讯作者：
David, Gregory