Covington M F, Panda S, Liu X L, Strayer C A, Wagner D R, Kay S A
Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403, USA.
Plant Cell. 2001 Jun;13(6):1305-15. doi: 10.1105/tpc.13.6.1305.
The Arabidopsis early flowering 3 (elf3) mutation causes arrhythmic circadian output in continuous light, but there is some evidence of clock function in darkness. Here, we show conclusively that normal circadian function occurs with no alteration of period length in elf3 mutants in dark conditions and that the light-dependent arrhythmia observed in elf3 mutants is pleiotropic on multiple outputs normally expressed at different times of day. Plants overexpressing ELF3 have an increased period length in both constant blue and red light; furthermore, etiolated ELF3-overexpressing seedlings exhibit a decreased acute CAB2 response after a red light pulse, whereas the null mutant is hypersensitive to acute induction. This finding suggests that ELF3 negatively regulates light input to both the clock and its outputs. To determine whether ELF3's action is phase dependent, we examined clock resetting by using light pulses and constructed phase response curves. Absence of ELF3 activity causes a significant alteration of the phase response curve during the subjective night, and constitutive overexpression of ELF3 results in decreased sensitivity to the resetting stimulus, suggesting that ELF3 antagonizes light input to the clock during the night. The phase of ELF3 function correlates with its peak expression levels in the subjective night. ELF3 action, therefore, represents a mechanism by which the oscillator modulates light resetting.
拟南芥早花3(elf3)突变在持续光照下会导致昼夜节律输出紊乱,但有证据表明其在黑暗中仍具有生物钟功能。在此,我们确凿地表明,在黑暗条件下,elf3突变体的昼夜节律功能正常,周期长度没有改变,并且在elf3突变体中观察到的光依赖性心律失常对通常在一天不同时间表达的多种输出具有多效性。过表达ELF3的植物在恒定蓝光和红光下周期长度均增加;此外,黄化的过表达ELF3的幼苗在红光脉冲后急性CAB2反应降低,而缺失突变体对急性诱导高度敏感。这一发现表明,ELF3对生物钟及其输出的光输入具有负调控作用。为了确定ELF3的作用是否依赖相位,我们通过光脉冲检查了生物钟重置并构建了相位响应曲线。ELF3活性的缺失会导致主观夜间相位响应曲线发生显著改变,而ELF3的组成型过表达会导致对重置刺激的敏感性降低,这表明ELF3在夜间拮抗光对生物钟的输入。ELF3功能的相位与其在主观夜间的峰值表达水平相关。因此,ELF3的作用代表了一种振荡器调节光重置的机制。