Domanski Michal, LaCava John
Department of Chemistry and Biochemistry, University of Bern, Bern, Switzerland.
Laboratory of Cellular and Structural Biology, the Rockefeller University, New York, USA.
Bio Protoc. 2017 Apr 20;7(8). doi: 10.21769/BioProtoc.2239.
The RNA exosome complex plays a central role in RNA processing and regulated turnover. Present both in cytoplasm and nucleus, the exosome functions through associations with ribonucleases and various adapter proteins (reviewed in [Kilchert ., 2016]). The RNA exosome-associated EXOSC10 protein is a distributive, 3'-5' exoribonuclease. The following protocol describes an approach to monitor the ribonucleolytic activity of affinity-purified EXOSC10-containing RNA exosomes, originating from HEK-293 cells, as reported in (Domanski ., 2016) and further detailed in the companion bio-protocol to this one (Domanski and LaCava, 2017).
RNA外切体复合物在RNA加工和调控的周转过程中起着核心作用。外切体存在于细胞质和细胞核中，通过与核糖核酸酶和各种衔接蛋白结合发挥作用（见[Kilchert ., 2016]综述）。与RNA外切体相关的EXOSC10蛋白是一种分布型的3'-5'外切核糖核酸酶。如下方案描述了一种监测源自HEK-293细胞的亲和纯化的含EXOSC10的RNA外切体核糖核酸酶活性的方法，如（Domanski ., 2016）所述，并在本方案的配套生物协议（Domanski和LaCava，2017）中有更详细的说明。
