The human origin recognition complex is essential for pre-RC assembly, mitosis, and maintenance of nuclear structure.
The human origin recognition complex is essential for pre-RC assembly, mitosis, and maintenance of nuclear structure.
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DOI：
10.7554/elife.61797
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发表时间：
2021-02-01
期刊：
影响因子：
7.7
通讯作者：
Stillman B
中科院分区：
文献类型：
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作者：
Chou HC;Bhalla K;Demerdesh OE;Klingbeil O;Hanington K;Aganezov S;Andrews P;Alsudani H;Chang K;Vakoc CR;Schatz MC;McCombie WR;Stillman B
起始点识别复合物（ORC）与CDC 6、MCM 2 -7和CDT 1协同作用，在DNA复制起始点形成前RC复合物。在这里，使用tiling-sgRNA CRISPR筛选，我们报告了ORC和CDC 6的每个亚基在人类细胞中是必不可少的。使用生长素诱导的降解系统，我们创建了能够快速消融ORC 2的稳定细胞系，揭示了多种细胞分裂周期表型。在缺乏ORC 2的情况下的主要缺陷是细胞在启动DNA复制或通过细胞分裂周期进展时遇到困难，这是由于G1期染色质上的MCM 2 -7负载减少。ORC 2缺陷细胞的细胞核也很大，异染色质不致密。一些ORC 2缺陷细胞完成DNA复制，进入有丝分裂，但从未退出。ORC 1敲除细胞也表现出极慢的细胞增殖和异常的细胞和核形态。因此，ORC蛋白和CDC 6对于正常细胞增殖是必不可少的，并有助于核组织。
The origin recognition complex (ORC) cooperates with CDC6, MCM2-7, and CDT1 to form pre-RC complexes at origins of DNA replication. Here, using tiling-sgRNA CRISPR screens, we report that each subunit of ORC and CDC6 is essential in human cells. Using an auxin-inducible degradation system, we created stable cell lines capable of ablating ORC2 rapidly, revealing multiple cell division cycle phenotypes. The primary defects in the absence of ORC2 were cells encountering difficulty in initiating DNA replication or progressing through the cell division cycle due to reduced MCM2-7 loading onto chromatin in G1 phase. The nuclei of ORC2-deficient cells were also large, with decompacted heterochromatin. Some ORC2-deficient cells that completed DNA replication entered into, but never exited mitosis. ORC1 knockout cells also demonstrated extremely slow cell proliferation and abnormal cell and nuclear morphology. Thus, ORC proteins and CDC6 are indispensable for normal cellular proliferation and contribute to nuclear organization.
DOI：
10.1083/jcb.142.2.307
发表时间：
1998-07-27
期刊：
The Journal of cell biology
影响因子：
--
作者：
Huang DW;Fanti L;Pak DT;Botchan MR;Pimpinelli S;Kellum R
通讯作者：
Kellum R