Overexpression of oncogenic H-Ras in hTERT-immortalized and SV40-transformed human cells targets replicative and specialized DNA polymerases for depletion.
Overexpression of oncogenic H-Ras in hTERT-immortalized and SV40-transformed human cells targets replicative and specialized DNA polymerases for depletion.
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DOI：
10.1371/journal.pone.0251188
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发表时间：
2021
期刊：
影响因子：
3.7
通讯作者：
Eckert KA
中科院分区：
文献类型：
--
作者：
Tsao WC;Buj R;Aird KM;Sidorova JM;Eckert KA
DNA聚合酶在复制叉进展和基因组维持中发挥重要作用。DNA损伤和药物诱导的复制应激导致特异性DNA聚合酶η和κ的上调和重新定位。虽然癌基因激活显着改变DNA复制动力学，导致复制应激和基因组的不稳定性，很少有人知道DNA聚合酶的表达和调节，在响应癌基因激活。在这里，我们研究了突变H-RAS G12 V过表达对h-TERT永生化和SV 40转化的人类细胞中DNA聚合酶调节的后果。专注于与复制叉相关的DNA聚合酶，我们证明了DNA聚合酶在响应H-RAS G12 V过表达的时间方式被耗尽。作为细胞显示衰老标志物的靶向消耗的聚合酶包括Pol α催化亚基（POLA 1）、Pol δ催化亚基和p68亚基（POLD 1和POLD 3）、Pol η和Pol κ。转录和转录后机制都介导这种反应。Pol η（POLH）耗竭足以诱导人包皮成纤维细胞BJ 5a细胞的衰老样生长停滞，并与Pol α表达降低相关。使用SV-40转化的细胞模型，我们观察到与Pol η缺陷细胞相比，H-RasG 12 V诱导的聚合酶耗尽的细胞中的细胞周期检查点信号传导差异。我们的研究结果有助于我们理解癌基因激活和细胞转化后的细胞事件。
DNA polymerases play essential functions in replication fork progression and genome maintenance. DNA lesions and drug-induced replication stress result in up-regulation and re-localization of specialized DNA polymerases η and κ. Although oncogene activation significantly alters DNA replication dynamics, causing replication stress and genome instability, little is known about DNA polymerase expression and regulation in response to oncogene activation. Here, we investigated the consequences of mutant H-RAS G12V overexpression on the regulation of DNA polymerases in h-TERT immortalized and SV40-transformed human cells. Focusing on DNA polymerases associated with the replication fork, we demonstrate that DNA polymerases are depleted in a temporal manner in response to H-RAS G12V overexpression. The polymerases targeted for depletion, as cells display markers of senescence, include the Pol α catalytic subunit (POLA1), Pol δ catalytic and p68 subunits (POLD1 and POLD3), Pol η, and Pol κ. Both transcriptional and post-transcriptional mechanisms mediate this response. Pol η (POLH) depletion is sufficient to induce a senescence-like growth arrest in human foreskin fibroblast BJ5a cells, and is associated with decreased Pol α expression. Using an SV-40 transformed cell model, we observed cell cycle checkpoint signaling differences in cells with H-RasG12V-induced polymerase depletion, as compared to Pol η-deficient cells. Our findings contribute to our understanding of cellular events following oncogene activation and cellular transformation.
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影响因子：
64.8
作者：
Bartkova, J;Horejsi, Z;Bartek, J
通讯作者：
Bartek, J
影响因子：
64.8
作者：
Bartkova, Jirina;Rezaei, Nousin;Gorgoulis, Vassilis G.
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Gorgoulis, Vassilis G.
影响因子：
64.8
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通讯作者：
di Fagagna, Fabrizio d'Adda
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8.8
作者：
Garcia-Exposito L;Bournique E;Bergoglio V;Bose A;Barroso-Gonzalez J;Zhang S;Roncaioli JL;Lee M;Wallace CT;Watkins SC;Opresko PL;Hoffmann JS;O'Sullivan RJ
通讯作者：
O'Sullivan RJ
影响因子：
11.4
作者：
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通讯作者：
Hoffmann, Jean-Sebastien