Department of Chemistry and Biochemistry, University of Colorado, Boulder, CO, USA.
Department of Chemistry and Biochemistry, University of Colorado, Boulder, CO, USA; Howard Hughes Medical Institute, University of Colorado, Boulder, CO, USA.
Methods. 2017 Aug 15;126:12-17. doi: 10.1016/j.ymeth.2017.04.020. Epub 2017 Apr 27.
Stress granules are dynamic, conserved RNA-protein (RNP) assemblies that form when translation is limiting; and are related to pathological aggregates in degenerative disease. Mammalian stress granules are comprised of two structures - an unstable shell and more stable cores. Herein we describe methodology for isolation of stress granule cores from both yeast and mammalian cells. The protocol consists of first enriching for stress granule cores using centrifugation and then further purifying stress granule cores using immunoprecipitation. The stress granule core isolation protocol provides a starting point for assisting future endeavors aimed at discovering conserved RNA regulatory mechanisms and potential links between RNP aggregation and degenerative disease.
应激颗粒是一种动态的、保守的 RNA-蛋白质（RNP）复合物，当翻译受到限制时会形成；与退行性疾病中的病理性聚集物有关。哺乳动物应激颗粒由两种结构组成 - 不稳定的外壳和更稳定的核心。在此，我们描述了从酵母和哺乳动物细胞中分离应激颗粒核心的方法。该方案首先使用离心法富集应激颗粒核心，然后使用免疫沉淀进一步纯化应激颗粒核心。应激颗粒核心分离方案为发现保守的 RNA 调节机制和 RNP 聚集与退行性疾病之间的潜在联系提供了一个起点。
