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In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity
Published on: January 29, 2018
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USP36 SUMOylates Las1L 并促进其在前核糖体RNA ITS2 处理中的功能
Yanping Li1, Yunhan Yang1, Rosalie C Sears1
1Department of Molecular and Medical Genetics, School of Medicine, and the OHSU Knight Cancer Institute, Oregon Health & Science University, Portland, Oregon.
Cancer research communications
|October 2, 2024
概括
无处不在特异性蛋白酶USP36通过控制Las1L-Nol9复合体来调节核糖体生物发生. 美国36美金
科学领域:
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
- 生物化学 生物化学
背景情况:
- 核糖体生物生成对细胞功能至关重要,其失调与癌症等疾病有关.
- Las1L-Nol9复合体对于60S核糖体成熟至关重要,特别是用于处理rRNA的内部转录间隔器2 (ITS2).
- 控制细胞内Las1L-Nol9复合体活动的精确调节机制在很大程度上是未知的.
研究的目的:
- 确定Las1L-Nol9复合体的新型调节体,并阐明它们在核糖体生物发生中的作用.
- 在Las1L-Nol9复合体和rRNA处理的背景下研究USP36的功能.
- 确定USP36影响Las1L-Nol9活动和核糖体生产的特定分子机制.
主要方法:
- 同免疫沉试验证实了USP36,Las1L和Nol9.9之间的相互作用.
- 西方涂抹以评估Las1L.的稳定性和修饰状态 (无化,SUMOylation).
- 针对Las1L的局部导向突变发生 (K565R) 为了研究SUMOylation的功能意义.
- 在内源Las1L被淘汰后进行功能性救援实验,以评估USP36和Las1L突变物在ITS2处理中的作用.
主要成果:
- USP36直接与Las1L和Nol9相互作用,通过duebiquitination调节它们的稳定性.
- USP36调解了Las1L的SUMOylation,主要是在素565 (K565) 中.
- 在ITS2处理中,K565转变为氨酸 (K565R) 废除了Las1L的功能,尽管保持了复杂的形成和蛋白质水平.
- 在K565中USP36介导的Las1L SUMOylation对于高效的ITS2处理和随后的核糖体生物生成至关重要.
结论:
- USP36作为一种新型的双重功能调节器,可以对Las1L进行脱氧化和SUMOylating.
- 在K565中通过USP36介导的Las1L的SUMOylation对于rRNA前ITS2处理至关重要.
- 这项研究揭示了 ribozom 生物发生的新调节途径,涉及 USP36 和 Las1L-Nol9 综合体.
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