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Cryo-Electron Microscopy of Endogenous Yeast Exosomes

  • Jun-Jie LiuEmail author
  • Hong-Wei WangEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2062)

Abstract

The RNA exosome is a multisubunit complex typically composed of a catalytically inactive core and the Rrp44 protein, which contains 3′-to-5′ exo- and endo-RNase activities. With assistance from nuclear or cytoplasmic cofactors, functional studies implicated the exosome as a critical player in the turnover of almost all RNA species, including mRNAs, rRNA, tRNAs, and other noncoding RNAs. Here, we describe the purification of the yeast 10-subunit exosome and 11-subunit Exosome-Ski7, as well as subsequent sample screening by negative staining EM and structural analysis by cryo-EM.

Key words

Exosome Rrp44 Ski7 RNA processing Cryo-EM 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Molecular Biophysics and Integrated Bioimaging DivisionLawrence Berkeley National LaboratoryBerkeleyUSA
  2. 2.Department of Molecular and Cell BiologyUniversity of California, BerkeleyBerkeleyUSA
  3. 3.Ministry of Education Key Laboratory of Protein Sciences, Tsinghua-Peking Joint Center for Life Sciences, Center for Structural Biology, School of Life SciencesTsinghua UniversityBeijingChina

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