mTOR-AKT signaling in cellular clock resetting triggered by osmotic stress.
mTOR-AKT signaling in cellular clock resetting triggered by osmotic stress.
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DOI：
10.1089/ars.2021.0059
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发表时间：
2022-01
影响因子：
6.6
通讯作者：
H. Yoshitane;Kiyomichi Imamura;Takenori Okubo;Yutaka Otobe;Satoshi Kawakami;Shunsuke Ito;Toru Takumi;K. Hattori;I. Naguro;H. Ichijo;Y. Fukada
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作者：
H. Yoshitane;Kiyomichi Imamura;Takenori Okubo;Yutaka Otobe;Satoshi Kawakami;Shunsuke Ito;Toru Takumi;K. Hattori;I. Naguro;H. Ichijo;Y. Fukada
目的 生物钟以细胞自主的方式振荡，周期约为 24 小时，相位通过多个时钟输入路径由光和温度等各种时间线索调节。我们之前发现渗透压和氧化应激强烈影响昼夜节律和细胞节律的相位，并且细胞凋亡信号调节激酶（ASK）家族成员Ask1、Ask2和Ask3的三重敲除消除了高渗脉冲治疗引起的相移（时钟重置）。我们的目的是探索依赖于 ASK 激酶的时钟输入通路中的关键分子和信号事件。结果 高渗脉冲治疗引起的细胞时钟相移因时钟（相关）基因 Dec1、Dec2 和 E4bp4 的联合缺陷而显着减少。此外，基于 LC-MS/MS 的蛋白质组学分析发现高渗脉冲诱导的 CLOCK Ser845 以 AKT 依赖性方式磷酸化。我们发现 AKT 激酶响应高渗脉冲实验而在 Ser473 位点被磷酸化（即激活）。 Torin 1 处理对 mTOR 激酶的抑制完全消除了 AKT 激活，抑制了 CLOCK Ser845 的磷酸化，并阻止了高渗脉冲处理诱导的时钟重置。结论我们得出结论，mTOR-AKT 信号对于 CLOCK Ser845 磷酸化是不可或缺的，因此会引发高渗脉冲治疗诱导的时钟重置。时钟（相关）基因和包含 Ser845 的 CLOCK C 末端区域的立即早期诱导也在通过氧化还原敏感的 ASK 激酶的时钟输入途径中发挥重要作用。
AIMS The circadian clock oscillates in a cell-autonomous manner with a period of approximately 24 hours, and the phase is regulated by various time cues such as light and temperature through multiple clock input pathways. We previously found that osmotic and oxidative stress strongly affected the circadian period and phase of cellular rhythms, and triple knock-out of apoptosis signal-regulating kinase (ASK) family members, Ask1, Ask2, and Ask3, abolished the phase shift (clock resetting) induced by hyperosmotic pulse treatment. We aimed at exploring a key molecule(s) and signaling events in the clock input pathway dependent on ASK kinases. RESULTS The phase shift of the cellular clock induced by the hyperosmotic pulse treatment was significantly reduced by combined deficiencies of the clock(-related) genes, Dec1, Dec2, and E4bp4. In addition, LC-MS/MS-based proteomic analysis identified hyperosmotic pulse-induced phosphorylation of CLOCK Ser845 in an AKT-dependent manner. We found that AKT kinase was phosphorylated at Ser473 (i.e., activated) in response to the hyperosmotic pulse experiments. Inhibition of mTOR kinase by Torin 1 treatment completely abolished the AKT activation, suppressed the phosphorylation of CLOCK Ser845, and blocked the clock resetting induced by the hyperosmotic pulse treatment. CONCLUSIONS We conclude that mTOR-AKT signaling is indispensable for the CLOCK Ser845 phosphorylation and consequently elicits the clock resetting induced by the hyperosmotic pulse treatment. Immediate early induction of the clock(-related) genes and CLOCK C-terminal region containing Ser845 also play important roles in the clock input pathway through redox-sensitive ASK kinases.