Reconstitution of circadian oscillation of cyanobacterial KaiC phosphorylation in vitro
Reconstitution of circadian oscillation of cyanobacterial KaiC phosphorylation in vitro
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DOI：
10.1126/science.1108451
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发表时间：
2005-04-15
期刊：
影响因子：
56.9
通讯作者：
Kondo, T
中科院分区：
文献类型：
--
作者：
Nakajima, M;Imai, K;Kondo, T
Kai蛋白全面调控蓝藻的昼夜基因表达。KaiC磷酸化周期，即使没有转录或翻译也会持续存在，被认为是生物钟的基本计时过程。我们通过将KaiC与KaiA、KaiB和三磷酸腺苷一起孵育，在体外重建了KaiC磷酸化的自我维持振荡。尽管温度变化（温度补偿），体外振荡的周期是稳定的，并且在体内KaiC突变株中观察到的昼夜节律周期与体外测量的昼夜节律周期一致。生物钟之谜现在可以在体外通过检查三种Kai蛋白之间的相互作用来研究。
Kai proteins globally regulate circadian gene expression of cyanobacteria. The KaiC phosphorylation cycle, which persists even without transcription or translation, is assumed to be a basic timing process of the circadian clock. We have reconstituted the self-sustainable oscillation of KaiC phosphorylation in vitro by incubating KaiC with KaiA, KaiB, and adenosine triphosphate. The period of the in vitro oscillation was stable despite temperature change (temperature compensation), and the circadian periods observed in vivo in KaiC mutant strains were consistent with those measured in vitro. The enigma of the circadian clock can now be studied in vitro by examining the interactions between three Kai proteins.