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The Eukaryotic RNA Exosome pp 427–448Cite as

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

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Part of the book series: Methods in Molecular Biology ((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.

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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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Authors and Affiliations

  1. Tri-Institutional Training Program in Chemical Biology, Memorial Sloan Kettering Cancer Center, New York, NY, USA

    John C. Zinder

  2. Structural Biology Program, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center, New York, NY, USA

    John C. Zinder & Christopher D. Lima

  3. Howard Hughes Medical Institute, New York, NY, USA

    Christopher D. Lima

Authors
  1. John C. Zinder
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  2. Christopher D. Lima
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Corresponding author

Correspondence to Christopher D. Lima .

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Editors and Affiliations

  1. Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, NY, USA

    John LaCava

  2. Central European Institute of Technology (CEITEC), Masaryk University, Brno, Czech Republic

    Štěpánka Vaňáčová

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Zinder, J.C., Lima, C.D. (2020). Reconstitution of S. cerevisiae RNA Exosome Complexes Using Recombinantly Expressed Proteins. In: LaCava, J., Vaňáčová, Š. (eds) The Eukaryotic RNA Exosome. Methods in Molecular Biology, vol 2062. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9822-7_21

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  • DOI: https://doi.org/10.1007/978-1-4939-9822-7_21

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9821-0

  • Online ISBN: 978-1-4939-9822-7

  • eBook Packages: Springer Protocols

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