生物
磷酸化
生物钟
激活剂(遗传学)
生物物理学
细胞生物学
生物化学
磷酸酶
受体
基因
作者
Yuta Shinohara,Yohei M. Koyama,Maki Ukai-Tadenuma,Takatsugu Hirokawa,Masaki Kikuchi,Rikuhiro G. Yamada,Hideki Ukai,Hiroshi Fujishima,Takashi Umehara,Kazuki Tainaka,Hiroki R. Ueda
出处
期刊:Molecular Cell
[Elsevier]
日期:2017-09-07
卷期号:67 (5): 783-798.e20
被引量:59
标识
DOI:10.1016/j.molcel.2017.08.009
摘要
Temperature compensation is a striking feature of the circadian clock. Here we investigate biochemical mechanisms underlying temperature-compensated, CKIδ-dependent multi-site phosphorylation in mammals. We identify two mechanisms for temperature-insensitive phosphorylation at higher temperature: lower substrate affinity to CKIδ-ATP complex and higher product affinity to CKIδ-ADP complex. Inhibitor screening of ADP-dependent phosphatase activity of CKIδ identified aurintricarboxylic acid (ATA) as a temperature-sensitive kinase activator. Docking simulation of ATA and mutagenesis experiment revealed K224D/K224E mutations in CKIδ that impaired product binding and temperature-compensated primed phosphorylation. Importantly, K224D mutation shortens behavioral circadian rhythms and changes the temperature dependency of SCN's circadian period. Interestingly, temperature-compensated phosphorylation was evolutionary conserved in yeast. Molecular dynamics simulation and X-ray crystallography demonstrate that an evolutionally conserved CKI-specific domain around K224 can provide a structural basis for temperature-sensitive substrate and product binding. Surprisingly, this domain can confer temperature compensation on a temperature-sensitive TTBK1. These findings suggest the temperature-sensitive substrate- and product-binding mechanisms underlie temperature compensation.
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