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Attenuation of the posttranslational oscillator via transcription–translation feedback enhances circadian-phase shifts in Synechococcus

机译:通过转录-翻译反馈对翻译后振荡器的衰减增强了Synechococcus中的昼夜节律相移。

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摘要

Circadian rhythms are endogenous biological timing processes that are ubiquitous in organisms ranging from cyanobacteria to humans. In the photoautotrophic unicellular cyanobacterium Synechococcus elongatus PCC 7942, under continuous light (LL) conditions, the transcription–translation feedback loop (TTFL) of KaiC generates a rhythmic change in the accumulation of KaiC relative to KaiA clock proteins (KaiC/KaiA ratio), which peak and trough at subjective dawn and dusk, respectively. However, the role of TTFL in the cyanobacterial circadian system remains unclear because it is not an essential requirement for the basic oscillation driven by the Kai-based posttranslational oscillator (PTO) and the transcriptional output mechanisms. Here, we show that TTFL is important for the circadian photic resetting property in Synechococcus. The robustness of PTO, which is exemplified by the amplitude of the KaiC phosphorylation cycle, changed depending on the KaiC/KaiA ratio, which was cyclic under LL. After cells were transferred from LL to the dark, the clock protein levels remained constant in the dark. When cells were transferred from LL to continuous dark at subjective dawn, the KaiC phosphorylation cycle was attenuated with a lower KaiC/KaiA ratio, a higher KaiC phosphorylation level, and a lower amplitude than that in cells transferred at subjective dusk. We also found that the greater the degree to which PTO was attenuated in continuous dark, the greater the phase shifts upon the subsequent light exposure. Based on these results, we propose that TTFL enhances resetting of the Kai-based PTO in Synechococcus.
机译:昼夜节律是内在的生物定时过程,在从蓝细菌到人类的生物中普遍存在。在光合自养单细胞蓝藻长突藻球藻PCC 7942中,在连续光照(LL)条件下,KaiC的转录-翻译反馈环(TTFL)相对于KaiA时钟蛋白(KaiC / KaiA比)产生KaiC积累的节律变化,分别在主观黎明和黄昏时达到高峰和低谷。但是,TTFL在蓝细菌昼夜节律系统中的作用仍不清楚,因为它不是由基于Kai的翻译后振荡器(PTO)驱动的基本振荡和转录输出机制的基本要求。在这里,我们表明TTFL对于Synechococcus中的昼夜节律复位特性很重要。以KaiC磷酸化周期的幅度为例的PTO的鲁棒性取决于在LL下呈环状的KaiC / KaiA比值。细胞从LL转移到黑暗后,时钟蛋白水平在黑暗中保持恒定。当在主观黎明时将细胞从LL转移到连续的黑暗时,与在主观黄昏转移的细胞相比,KaiC / KaiA比降低,KaiC磷酸化水平更高和幅度更低时,KaiC磷酸化周期被减弱。我们还发现,在连续的黑暗中,PTO衰减的程度越大,随后的曝光时相移就越大。基于这些结果,我们建议TTFL增强Synechococcus中基于Kai的PTO的复位。

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