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Updated: Jan 9, 2026

Cycloheximide Chase Analysis of Protein Degradation in Saccharomyces cerevisiae

Cycloheximide Chase Analysis of Protein Degradation in Saccharomyces cerevisiae

Published on: April 18, 2016

在Saccharomyces cerevisiae中异常的凝聚功能激活Mcd1降解以促进细胞致死性

Gurvir Singh¹, Robert V Skibbens¹

¹Department of Biological Sciences, Lehigh University, 111 Research Drive, Bethlehem, Pennsylvania, 18015. United States of America.

bioRxiv : the preprint server for biology

|December 4, 2025

概括

通过E3结合酶驱动的Mcd1蛋白降解驱动的凝聚素复合体失活,促进细胞致死性. 这种降解起到负反机制的作用,在S阶段维持Mcd1的稳态.

科学领域:

细胞生物学细胞生物学
分子遗传学分子遗传学
基因组学就是基因组学.

背景情况:

包含Smc1,Smc3和Mcd1蛋白质的凝聚蛋白复合体对于DNA复制,修复和基因转录等过程至关重要.
凝聚素的功能与细胞周期检查点有关,但这些检查点如何应对凝聚素功能障碍尚不清楚.

研究的目的:

为了研究凝聚素突变体中Mcd1蛋白质损失的机制.
要确定Mcd1损失是否是凝聚力不稳定的原因或后果.
为了确定调节Mcd1水平的细胞通路,以应对异常的凝聚功能.

主要方法:

在各种凝聚蛋白突变体中分析Mcd1蛋白水平 (rad61,scc2-4).
评估Mcd1重新表达对温度敏感凝聚性突变细胞生长的影响.
使用特定突变物 (例如,San1) 调查E3酶参与Mcd1降解.
在细胞周期 (S阶段) 中监测Mcd1水平和转录.

主要成果:

麦克德1的损失独立于凝聚素复合体的完整性发生,即使在具有稳定凝聚素的突变物中也存在.
恢复Mcd1水平在所有测试的凝聚素等位基因中挽救了生长缺陷,这表明Mcd1损失对失活化至关重要.
异常的凝聚功能触发了Mcd1通过San1.1等E3酶的降解.
在S阶段,Mcd1被降解,抵消转录增加和典型的峰值水平,这表明反循环.

结论:

Mcd1蛋白质的损失是凝聚素复合体失活的关键机制,导致细胞致死性.
细胞利用E3酶来积极降解Mcd1,以应对凝聚素功能障碍.
负反机制调节了Mcd1的稳态,特别是在S阶段,尽管转录上调.

关键词:

检查站检查站检查站凝聚素是一种合素.康尼利亚·德朗格综合征 (CdLS)这就是 Das1 Das1 Das1 E3 结合酶的使用在Eco1/Ctf7/ESCO2中使用.在MCD1 / SCC1 / RAD21 中,MCD1 / SCC1 / RAD21 在MCD1 / SCC1 / RAD21 中.在Rad61/WAPL中使用.罗伯茨综合症 (RBS) 是一种卫生院卫生院染色体凝聚凝聚的情况.姐妹染色体凝聚力的一致性

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