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Reconstitution of S. cerevisiae RNA Exosome Complexes Using Recombinantly Expressed Proteins

  • John C. Zinder
  • Christopher D. LimaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2062)

Abstract

3′ to 5′ RNA degradation is primarily catalyzed by the RNA exosome subunits Dis3 and Rrp6 in the nucleus of Saccharomyces cerevisiae. These enzymes form a complex with the nine-subunit noncatalytic core (Exo9) to carry out their functions in vivo. Protein cofactors Rrp47, Mpp6, and the Mtr4 RNA helicase also assist the complex by modulating its activities and/or recruiting it to specific RNAs for processing or degradation. Here we present our preferred strategy for reconstituting RNA exosomes from S. cerevisiae using purified, recombinantly expressed components.

Key words

RNA exosome RNA decay Exoribonuclease Budding yeast 

Notes

Acknowledgments

This work was supported in part by GM065872 and GM118080 (NIH/NIGMS, C.D.L) and P30CA008748 (NIH/National Cancer Institute). The content is the authors’ responsibility and does not represent the official views of the NIH. C.D.L is a Howard Hughes Medical Institute Investigator.

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Copyright information

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

Authors and Affiliations

  1. 1.Tri-Institutional Training Program in Chemical BiologyMemorial Sloan Kettering Cancer CenterNew YorkUSA
  2. 2.Structural Biology Program, Sloan Kettering InstituteMemorial Sloan Kettering Cancer CenterNew YorkUSA
  3. 3.Howard Hughes Medical InstituteNew YorkUSA

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