The non-catalytic role of DNA polymerase epsilon in replication initiation in human cells.
The non-catalytic role of DNA polymerase epsilon in replication initiation in human cells.
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DOI：
10.1038/s41467-022-34911-4
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发表时间：
2022-11-19
影响因子：
16.6
通讯作者：
Moiseeva, Tatiana N.
中科院分区：
文献类型：
--
作者：
Vipat, Sameera;Gupta, Dipika;Jonchhe, Sagun;Anderspuk, Hele;Rothenberg, Eli;Moiseeva, Tatiana N.
DNA聚合酶抑制剂（PolE）是DNA复制所必需的酶。PolE的缺陷和突变会导致严重的发育异常和癌症。特别地，酵母Pole催化亚基的催化结构域是存活的关键，其非催化的基本功能与复制解旋酶（CMG）组装有关。关于PolE在人类细胞复制起始中的作用知之甚少。在这里，我们使用一个生长素诱导的降解决定子系统来研究POLE 1缺失对U2 OS细胞复制起始的影响。POLE 1缺失的细胞能够组装CMG解旋酶并启动DNA合成，但不久后就失败了。POLE 1非催化结构域的表达挽救了这种缺陷，导致缓慢但连续的DNA合成。我们提出了一个模型，其中在人类U2 OS细胞POLE 1/POLE 2是CMG组装的关键，但在复制起始的后期步骤中是必不可少的。我们的研究为PolE在人类细胞复制起始中的作用提供了一些见解。DNA聚合酶β在DNA复制起始中具有关键作用。在这里，作者表明，在人类癌细胞中，POLE是复制解旋酶组装，但不是复制起始，这需要POLE 1的非催化结构域。
DNA polymerase epsilon (PolE) in an enzyme essential for DNA replication. Deficiencies and mutations in PolE cause severe developmental abnormalities and cancers. Paradoxically, the catalytic domain of yeast PolE catalytic subunit is dispensable for survival, and its non-catalytic essential function is linked with replicative helicase (CMG) assembly. Less is known about the PolE role in replication initiation in human cells. Here we use an auxin-inducible degron system to study the effect of POLE1 depletion on replication initiation in U2OS cells. POLE1-depleted cells were able to assemble CMG helicase and initiate DNA synthesis that failed shortly after. Expression of POLE1 non-catalytic domain rescued this defect resulting in slow, but continuous DNA synthesis. We propose a model where in human U2OS cells POLE1/POLE2 are dispensable for CMG assembly, but essential during later steps of replication initiation. Our study provides some insights into the role of PolE in replication initiation in human cells. DNA polymerase epsilon has a critical role in DNA replication initiation. Here, the authors show that in human cancer cells POLE is dispensable for the replicative helicase assembly but not for replication initiation, which requires the non-catalytic domain of POLE1.
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DOI：
10.15252/embj.2021108819
发表时间：
2021-12-01
期刊：
The EMBO journal
影响因子：
--
作者：
Jones ML;Baris Y;Taylor MRG;Yeeles JTP
通讯作者：
Yeeles JTP
影响因子：
14.9
作者：
Ma J;Chen T;Wu S;Yang C;Bai M;Shu K;Li K;Zhang G;Jin Z;He F;Hermjakob H;Zhu Y
通讯作者：
Zhu Y
影响因子：
16.6
作者：
Garbacz MA;Lujan SA;Burkholder AB;Cox PB;Wu Q;Zhou ZX;Haber JE;Kunkel TA
通讯作者：
Kunkel TA
影响因子：
14.9
作者：
Gödecke N;Zha L;Spencer S;Behme S;Riemer P;Rehli M;Hauser H;Wirth D
通讯作者：
Wirth D
影响因子：
16
作者：
Bellelli R;Borel V;Logan C;Svendsen J;Cox DE;Nye E;Metcalfe K;O'Connell SM;Stamp G;Flynn HR;Snijders AP;Lassailly F;Jackson A;Boulton SJ
通讯作者：
Boulton SJ