Rutila J E, Zeng H, Le M, Curtin K D, Hall J C, Rosbash M
Howard Hughes Medical Institute, National Science Foundation, Science and Technology Center for Biological Timing.
Neuron. 1996 Nov;17(5):921-9. doi: 10.1016/s0896-6273(00)80223-8.
To identify new components of the Drosophila circadian clock, we screened chemically mutagenized flies for suppressors or enhancers of the long periods characteristic of the period (per) mutant allele perL. We isolated a novel mutant that maps to the rhythm gene timeless (tim). This novel allele, timSL, alters the temporal pattern of perL protein nuclear localization and restores temperature compensation to perL flies. timSL more generally manifests specific interactions with different per alleles. The identification of this first period-altering tim allele provides further evidence that TIM is a major component of the clock, and the allele-specific interactions with PER provide evidence that the PER/TIM heterodimer is a unit of circadian function. Although timSL fails to restore PER-L/TIM temperature insensitivity in yeast, it alters the TIM phosphorylation pattern during the late night. The effects on phosphorylation suggest that timSL functions as a partial bypass suppressor of perL and provide evidence that the TIM phosphorylation program contributes to the circadian timekeeping mechanism.
为了鉴定果蝇生物钟的新组分,我们对经化学诱变的果蝇进行了筛选,寻找周期(per)突变等位基因perL所具有的长周期特征的抑制子或增强子。我们分离出一个新的突变体,它定位于节律基因无时间(tim)。这个新的等位基因timSL改变了perL蛋白核定位的时间模式,并恢复了perL果蝇的温度补偿。timSL更普遍地表现出与不同per等位基因的特异性相互作用。这个首个改变周期的tim等位基因的鉴定进一步证明TIM是生物钟的主要组分,并且与PER的等位基因特异性相互作用证明PER/TIM异二聚体是昼夜节律功能的一个单元。尽管timSL未能在酵母中恢复PER-L/TIM的温度不敏感性,但它在深夜改变了TIM的磷酸化模式。对磷酸化的影响表明timSL作为perL的部分旁路抑制子发挥作用,并证明TIM磷酸化程序有助于昼夜节律计时机制。