Reconstitution, activities, and structure of the eukaryotic RNA exosome
Reconstitution, activities, and structure of the eukaryotic RNA exosome
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DOI：
10.1016/j.cell.2006.10.037
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发表时间：
2006-12-15
期刊：
影响因子：
64.5
通讯作者：
Lima, Christopher D.
中科院分区：
文献类型：
--
作者：
Liu, Quansheng;Greimann, Jaclyn C.;Lima, Christopher D.
RNA外泌体是一种多亚基3'至5'核糖核酸外切酶复合物，参与细胞RNA的降解和加工。为了确定真核外泌体的活性和结构，我们报告了来自酵母和人的9亚基外泌体的重建以及来自酵母的10亚基和11亚基外泌体的重建。使用富含AU的、多腺苷酸化（poly[A]）的、通用的和结构化的RNA底物对纯化的亚基和重构的外泌体进行的比较生物化学分析揭示了人Rrp 41/Rrp 45和9亚基人外泌体的进行性磷酸化活性、酵母Rrp 44和酵母10亚基外泌体的进行性水解活性、Rrp 6的分配性水解活性、以及酵母11-亚基外泌体的进行性和分配性水解活性。为了阐明真核外泌体的结构，其保守的表面，和RNA衰变的结构基础，我们报告了在3.35埃的286 kDa的九亚基人类外泌体的X射线结构测定。
The RNA exosome is a multisubunit 3' to 5' exoribonuclease complex that participates in degradation and processing of cellular RNA. To determine the activities and structure of the eukaryotic exosome, we report the reconstitution of 9-subunit exosomes from yeast and human and reconstitution of 10- and 11-subunit exosomes from yeast. Comparative biochemical analysis between purified subunits and reconstituted exosomes using AU-rich, polyadenylated (poly[A]), generic, and structured RNA substrates reveals processive phosphorolytic activities for human Rrp41/Rrp45 and the 9-subunit human exosome, processive hydrolytic activities for yeast Rrp44 and the yeast 10-sub-unit exosome, distributive hydrolytic activities for Rrp6, and processive and distributive hydrolytic activities for the yeast 11-subunit exosome. To elucidate the architecture of a eukaryotic exosome, its conserved surfaces, and the structural basis for RNA decay, we report the X-ray structure determination for the 286 kDa nine-subunit human exosome at 3.35 angstrom.