Rad9, Rad17, TopBP1 and claspin play essential roles in heat-induced activation of ATR kinase and heat tolerance.
Rad9, Rad17, TopBP1 and claspin play essential roles in heat-induced activation of ATR kinase and heat tolerance.
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DOI：
10.1371/journal.pone.0055361
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Maehara Y
中科院分区：
文献类型：
--
作者：
Tuul M;Kitao H;Iimori M;Matsuoka K;Kiyonari S;Saeki H;Oki E;Morita M;Maehara Y
热疗被广泛用于治疗癌症患者，特别是与其他治疗方法如放射治疗相结合。热处理本身激活由ATR-Chk 1和ATM-Chk 2途径介导的DNA损伤反应，但尚未完全理解这些DNA损伤反应是如何被激活并影响耐热性的。通过对人HeLa细胞和鸡B淋巴瘤DT 40细胞进行遗传分析，我们发现ATR诱导的热诱导Chk 1 Ser 345磷酸化在很大程度上依赖于Rad 9、Rad 17、TopBP 1和Claspin。然而，通过加热激活ATR-Chk 1通路与FancD 2单泛素化或RPA 32磷酸化无关，FancD 2单泛素化或RPA 32磷酸化是复制叉停滞时ATR激酶激活的下游事件。ATR、Rad 9、Rad 17、TopBP 1或Claspin的下调显著降低了热疗后的克隆形成细胞活力，而基因敲除或ATM激酶的抑制仅适度降低了克隆形成细胞活力。抑制ATR-Chk 1通路激活增强热诱导的Chk 2 Thr 68磷酸化，同时抑制ATR和ATM激酶，导致严重的热细胞毒性。这些数据表明，ATR-Chk 1途径在停滞的复制叉激活的必要因素也需要热诱导激活ATR激酶，这主要有助于耐热性在一个非重叠的方式与ATM激酶。
Hyperthermia is widely used to treat patients with cancer, especially in combination with other treatments such as radiation therapy. Heat treatment per se activates DNA damage responses mediated by the ATR-Chk1 and ATM-Chk2 pathways but it is not fully understood how these DNA damage responses are activated and affect heat tolerance. By performing a genetic analysis of human HeLa cells and chicken B lymphoma DT40 cells, we found that heat-induced Chk1 Ser345 phosphorylation by ATR was largely dependent on Rad9, Rad17, TopBP1 and Claspin. Activation of the ATR-Chk1 pathway by heat, however, was not associated with FancD2 monoubiquitination or RPA32 phosphorylation, which are known as downstream events of ATR kinase activation when replication forks are stalled. Downregulation of ATR, Rad9, Rad17, TopBP1 or Claspin drastically reduced clonogenic cell viability upon hyperthermia, while gene knockout or inhibition of ATM kinase reduced clonogenic viability only modestly. Suppression of the ATR-Chk1 pathway activation enhanced heat-induced phosphorylation of Chk2 Thr68 and simultaneous inhibition of ATR and ATM kinases rendered severe heat cytotoxicity. These data indicate that essential factors for activation of the ATR-Chk1 pathway at stalled replication forks are also required for heat-induced activation of ATR kinase, which predominantly contributes to heat tolerance in a non-overlapping manner with ATM kinase.
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影响因子：
5.3
作者：
Kim, JE;McAvoy, SA;Chen, JJ
通讯作者：
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影响因子：
9.2
作者：
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DOI：
10.1016/s0360-3016(00)01465-6
发表时间：
2001-04-01
影响因子：
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影响因子：
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影响因子：
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作者：
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