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Related Experiment Videos

Yeast Trf5p is a nuclear poly(A) polymerase.

Jonathan Houseley1, David Tollervey

  • 1Wellcome Trust Centre for Cell Biology, University of Edinburgh, Swann Building, King's Buildings, Edinburgh EH9 3JR, UK.

EMBO Reports
|December 24, 2005
PubMed
Summary

The yeast nuclear exosome

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Area of Science:

  • Molecular Biology
  • Yeast Genetics
  • RNA Metabolism

Background:

  • The yeast nuclear exosome degrades RNA.
  • The Trf4p-Air1/2p-Mtr4p (TRAMP) complex stimulates exosome activity.
  • Rrp6p is a key component of the nuclear exosome.

Purpose of the Study:

  • To investigate the role of Rrp6p in polyadenylation of ribosomal RNA precursors (pre-rRNAs).
  • To characterize the function of Trf5p and its relationship with the TRAMP complex.

Main Methods:

  • Analysis of yeast strains with deletions or overexpression of specific genes (Rrp6p, Trf4p, Trf5p, Mtr4p).
  • Polyadenylation assays to measure pre-rRNA modification.
  • In vitro polyadenylation assays using purified Trf5p.
  • Co-purification studies to identify protein complexes.

Main Results:

  • Strains lacking Rrp6p accumulate polyadenylated pre-rRNAs.
  • Polyadenylation is reduced by Trf4p or Trf5p deletion and enhanced by Trf5p overexpression.
  • Mtr4p is essential for coupling polyadenylation to degradation.
  • Trf5p exhibits in vitro polyadenylation activity and forms the TRAMP5 complex with Mtr4p and Air1p.

Conclusions:

  • Rrp6p is involved in regulating pre-rRNA polyadenylation.
  • The TRAMP5 complex, containing Trf5p, plays a role in pre-rRNA processing.
  • Mtr4p is crucial for the functional coordination of polyadenylation and RNA degradation pathways.

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