The Ataxia telangiectasia-mutated and Rad3-related protein kinase regulates cellular hydrogen sulfide concentrations
The Ataxia telangiectasia-mutated and Rad3-related protein kinase regulates cellular hydrogen sulfide concentrations
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DOI：
10.1016/j.dnarep.2018.11.002
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发表时间：
2019-01-01
期刊：
影响因子：
3.8
通讯作者：
Shackelford, Rodney E.
中科院分区：
文献类型：
--
作者：
Chen, Jie;Shen, Xinggui;Shackelford, Rodney E.
共济失调毛细血管扩张症突变和Rad 3相关（ATR）丝氨酸/苏氨酸激酶在修复复制相关的DNA损伤，维持S和G2/M期基因组稳定性以及促进有丝分裂染色体分离中起着重要作用。许多刺激激活ATR，包括在停滞复制的人中持续的单链DNA、R环形成、缺氧、紫外线和氧化应激，导致ATR介导的蛋白磷酸化。近年来，硫化氢（H2S）作为一种内源性气体递质，在类似的细胞应激环境下通过复杂的氧化还原反应调节多种细胞过程。三种酶合成H2S：胱硫醚-β-合酶、胱硫醚γ-裂解酶和3-巯基丙酮酸硫转移酶。由于H_2S在某些条件下可引起DNA损伤，我们推测ATR活性可能调节细胞H_2S浓度和H_2S合成酶。在这里，我们表明，携带双等位基因敲入亚型ATR突变的人结直肠癌细胞具有较低的细胞H2S浓度比同源ATR野生型细胞，和所有三个H2S合成酶显示较低的蛋白质表达在ATR亚型突变细胞。此外，ATR丝氨酸428磷酸化被H2S供体和H2S合成酶抑制改变，而氧化应激诱导的ATR调节蛋白CHK 1在丝氨酸345上的磷酸化被H2S合成酶抑制增加。最后，抑制H2S的产生增强氧化应激诱导的双链DNA断裂的ATR亚型突变体相比，ATR野生型细胞。我们的研究结果表明，ATR激酶的调节和H2S的调节。
The ataxia telangiectasia-mutated and Rad3-related (ATR) serine/threonine kinase plays a central role in the repair of replication-associated DNA damage, the maintenance of S and G2/M-phase genomic stability, and the promotion of faithful mitotic chromosomal segregation. A number of stimuli activate ATR, including persistent single-stranded DNA at stalled replication folks, R loop formation, hypoxia, ultraviolet light, and oxidative stress, leading to ATR-mediated protein phosphorylation. Recently, hydrogen sulfide (H2S), an endogenous gasotransmitter, has been found to regulate multiple cellular processes through complex redox reactions under similar cell stress environments. Three enzymes synthesize H2S: cystathionine-beta-synthase, cystathionine gamma-lyase, and 3-mercaptopyruvate sulfurtransferase. Since H2S can under some conditions cause DNA damage, we hypothesized that ATR activity may regulate cellular H2S concentrations and H2S-syntheszing enzymes. Here we show that human colorectal cancer cells carrying biallelic knock-in hypomorphic ATR mutations have lower cellular H2S concentrations than do syngeneic ATR wild-type cells, and all three H2S-synthesizing enzymes show lower protein expression in the ATR hypomorphic mutant cells. Additionally, ATR serine 428 phosphorylation is altered by H2S donor and H2S synthesis enzyme inhibition, while the oxidative-stress induced phosphorylation of the ATR-regulated protein CHK1 on serine 345 is increased by H2S synthesis enzyme inhibition. Lastly, inhibition of H2S production potentiated oxidative stress-induced double-stranded DNA breaks in the ATR hypomorphic mutant compared to ATR wild-type cells. Our findings demonstrate that the ATR kinase regulates and is regulated by H2S.