Sammelbandbeitrag

Erschienen in: The Eukaryotic RNA Exosome

Herausgeberliste: LaCava, J.; Vaňáčová, Š.

Jahr der Veröffentlichung: 2020

Seiten: 63-79

ISBN: 978-1-4939-9821-0

eISBN: 978-1-4939-9822-7

DOI Link: https://doi.org/10.1007/978-1-4939-9822-7_4

Serientitel: Methods in Molecular Biology

Serienzählung: 2062

Abstract: 

The archaeal exosome is a protein complex with phosphorolytic activity. It is built of a catalytically active hexameric ring containing the archaeal Rrp41 and Rrp42 proteins, and a heteromeric RNA-binding platform. The platform contains a heterotrimer of the archaeal Rrp4 and Csl4 proteins (which harbor S1 and KH or Zn-ribbon RNA binding domains), and comprises additional archaea-specific subunits. The latter are represented by the archaeal DnaG protein, which harbors a novel RNA-binding domain and tightly interacts with the majority of the exosome isoforms, and Nop5, known as a part of an rRNA methylating complex and found to associate with the archaeal exosome at late stationary phase. Although in the cell the archaeal exosome exists in different isoforms with heterotrimeric Rrp4-Csl4-caps, in vitro it is possible to reconstitute complexes with defined, homotrimeric caps and to study the impact of each RNA-binding subunit on exoribonucleolytic degradation and on polynucleotidylation of RNA. Here we describe procedures for reconstitution of isoforms of the Sulfolobus solfataricus exosome and for set-up of RNA degradation and polyadenylation assays.

Harvard-Zitierstil: Evguenieva-Hackenberg, E., Gauernack, A., Hou, L. and Klug, G. (2020) Enzymatic Analysis of Reconstituted Archaeal Exosomes, in LaCava, J. and Vaňáčová, Š. (eds.) The Eukaryotic RNA Exosome. New York, NY: Humana, pp. 63-79. https://doi.org/10.1007/978-1-4939-9822-7_4

APA-Zitierstil: Evguenieva-Hackenberg, E., Gauernack, A., Hou, L., & Klug, G. (2020). Enzymatic Analysis of Reconstituted Archaeal Exosomes. In LaCava, J., & Vaňáčová, Š. (Eds.), The Eukaryotic RNA Exosome (pp. 63-79). Humana. https://doi.org/10.1007/978-1-4939-9822-7_4