在裂变酵母中对内源蛋白进行化学遗传操纵,使用自我定位的带诱导蛋白转位系统
Akinobu Nakamura1, Yuhei Goto1,2,3, Hironori Sugiyama1
1Quantitative Biology Research Group, Exploratory Research Center on Life and Living Systems (ExCELLS), National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji-cho, Okazaki, Aichi 444-8787, Japan.
ACS chemical biology
|November 22, 2023
概括
研究人员开发了一种新的化学遗传工具,即自我定位联体诱导蛋白转位 (SLIPT) 系统,用于精确控制裂变酵母中的蛋白质定位. 该系统能够快速扰乱细胞周期蛋白质,为研究细胞信号提供更好的时间分辨率.
科学领域:
- 细胞生物学 细胞生物学
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
背景情况:
- 细胞利用细胞内信号网络来处理外部刺激,通常涉及蛋白质转位.
- 化学诱导二分化 (CID) 系统改变了蛋白质的局部化,但在裂变酵母中存在局限性,需要更好的工具.
- 现有的方法,如Schizosaccharomyces pombe中的FKBP-FRB,面临技术挑战,需要改进的扰动工具.
研究的目的:
- 应用和验证自局部联体诱导蛋白转位 (SLIPT) 系统在*Schizosaccharomyces pombe*.
- 评估SLIPT系统在扰乱细胞循环相关蛋白质和内源分子方面的有效性.
- 建立具有高时间分辨率的化学遗传工具,用于研究细胞内信号和细胞循环调节.
主要方法:
- 在S. pombe*中实施自我定位的联体诱导蛋白转位 (SLIPT) 系统.
- 优化自我定位的配体,以持续招募结合伙伴到细胞区,特别是等离子体膜.
- 对编码结合伙伴的基因进行内置,以研究内源分子的局部化和功能.
主要成果:
- 在S. pombe*中使用SLIPT系统成功扰乱了几个与细胞循环相关的蛋白质.
- 通过优化的自我定位连接物,证明了对分子的长期招募到等离子体膜.
- 观察到内源分子局部化的变化,导致细胞周期进展和相关表型的扰乱.
结论:
- SLIPT系统是一种有效的化学遗传工具,用于快速扰乱S. pombe*中的内源分子.
- 该系统提供了一种有价值的方法,用于研究具有增强时间分辨率的细胞内信号通路.
- 这项研究强调了SLIPT系统在促进酵母细胞循环调节和分子信号传导方面的研究潜力.
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