Transcription of the Neurospora crassa circadian clock gene frequency (frq) is an essential process in the negative feedback loop that controls circadian rhythms. WHITE COLLAR 1 (WC-1) and WHITE COLLAR 2 (WC-2) forms the WC complex (WCC) that is the main activator of frq transcription by binding to its promoter. Here, we show that Centromere Binding Factor 1 (CBF-1) is a critical component of the N. crassa circadian clock by regulating frq transcription. Deletion of cbf-1 resulted in long period and low amplitude rhythms, whereas overexpression of CBF-1 abolished the circadian rhythms. Loss of CBF-1 resulted in WC-independent FRQ expression and suppression of WCC activity. As WCC, CBF-1 also binds to the C-box at the frq promoter. Overexpression of CBF-1 impaired WCC binding to the C-box to suppress frq transcription. Together, our results suggest that the proper level of CBF-1 is critical for circadian clock function by suppressing WC-independent FRQ expression and by regulating WCC binding to the frq promoter.

译文

:Neurospora crassa昼夜节律时钟基因频率(frq)的转录是控制昼夜节律的负反馈回路中的重要过程。 WHITE COLLAR 1(WC-1)和WHITE COLLAR 2(WC-2)形成WC复合物(WCC),它是frq转录的主要激活因子,与它的启动子结合。在这里,我们显示通过调节frq转录,着丝粒结合因子1(CBF-1)是N. crassa昼夜节律时钟的关键组成部分。删除cbf-1会导致长周期和低振幅节律,而CBF-1的过表达则消除了昼夜节律。 CBF-1的丢失导致WC独立的FRQ表达和WCC活性的抑制。作为WCC,CBF-1在frq启动子上也与C-box结合。 CBF-1的过表达会损害WCC与C-box的结合,从而抑制frq转录。总之,我们的结果表明,通过抑制WC独立的FRQ表达并调节WCC与frq启动子的结合,CBF-1的适当水平对于昼夜节律功能至关重要。

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