Kim Dong-Hwan, Koepp Deanna M
Department of Genetics, Cell Biology and Development, University of Minnesota, 6-160 Jackson Hall, 321 Church Street SE, Minneapolis, MN, 55455, USA.
Methods Mol Biol. 2014;1170:343-56. doi: 10.1007/978-1-4939-0888-2_17.
Cell cycle progression is tightly regulated to prevent uncontrolled growth and division. Specific cell cycle factors are responsible for driving the cell from one cell cycle stage to the next. Many of these proteins are targeted for degradation by the ubiquitin proteasome system when their function is no longer required or may disrupt cell cycle progression. Here we describe a series of experiments used to study the ubiquitin-mediated degradation of cell cycle proteins. This collection of assays may be used to determine the requirement for individual proteins in the degradation and ubiquitination of cell cycle proteins in Saccharomyces cerevisiae.
细胞周期进程受到严格调控,以防止细胞不受控制地生长和分裂。特定的细胞周期因子负责推动细胞从一个细胞周期阶段进入下一个阶段。当这些蛋白质的功能不再需要或可能扰乱细胞周期进程时,它们中的许多会被泛素蛋白酶体系统靶向降解。在这里,我们描述了一系列用于研究细胞周期蛋白泛素介导降解的实验。这组分析方法可用于确定酿酒酵母中细胞周期蛋白降解和泛素化过程中对单个蛋白质的需求。