活的多索菲拉细胞中的PER-TIM相互作用:为昼夜时钟的间隔计时器
Pablo Meyer1, Lino Saez, Michael W Young
1Laboratory of Genetics, Rockefeller University, 1230 York Avenue, New York, NY 10021, USA.
Science (New York, N.Y.)
|January 18, 2006
概括
循环时钟蛋白PER和TIM在细胞质中迅速结合和积累,然后突然解离并进入细胞核. 这种时间调节不受每次L) 突变的影响,是时钟周期性的关键.
科学领域:
- 时间生物学 时间生物学
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
背景情况:
- 昼夜时钟依赖于核心时钟蛋白的循环表达和定位.
- 目前的模型表明,对于周期 (PER) 和无时无刻 (TIM) 蛋白质的局部化动态,有一个更简单的模型.
研究的目的:
- 为了研究PER和TIM蛋白相互作用和单细胞内定位的精确动态.
- 探索per(L) 突变在昼夜钟蛋白的时间调节中的作用.
主要方法:
- 使用光标记PER和TIM蛋白质的单细胞成像试验.
- 采用光共振能量转移 (FRET) 测量来监测蛋白质结合和相互作用.
- 在培养细胞和体内观察到蛋白质定位和动态.
主要成果:
- PER和TIM蛋白迅速结合并在细胞质中持续存在,形成离散的焦点.
- 复合体在大约6小时后突然解离,PER和TIM在狭窄的时间范围内独立进入核.
- L突变延迟了核进入,但没有改变PER/TIM组装或解离率.
结论:
- 揭示了一个以前未知的时间调节机制在昼夜时钟功能.
- 证明了核进入的时间,而不是组装或解离,对昼夜周期性至关重要.
- 强调了精确的蛋白质动态在维持昼夜节律的重要性.
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