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Analysis of Translation Initiation During Stress Conditions by Polysome Profiling
10:59

Analysis of Translation Initiation During Stress Conditions by Polysome Profiling

Published on: May 19, 2014

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The polysome-associated proteins Scp160 and Bfr1 prevent P body formation under normal growth conditions

Julie Weidner1, Congwei Wang, Cristina Prescianotto-Baschong

  • 1Growth & Development, Biozentrum, University of Basel, Klingelbergstrasse 70, 4056 Basel, Switzerland.

Journal of Cell Science
|February 27, 2014
PubMed

Insights

The mRNA-binding proteins Bfr1 and Scp160 normally prevent processing bodies (P bodies) from forming in yeast cells. Their absence triggers P body formation, even without cellular stress.

Area of Science:

  • Molecular Biology
  • Yeast Genetics
  • RNA Metabolism

Background:

  • Processing bodies (P bodies) are crucial cellular structures involved in mRNA storage and degradation in Saccharomyces cerevisiae.
  • Under normal growth conditions, P bodies are rarely observed, but their number increases significantly upon cellular stress.
  • The regulation of P body formation and its relationship with mRNA metabolism under non-stress conditions are not fully understood.

Purpose of the Study:

  • To investigate the role of the mRNA-binding protein Scp160 and its interaction partner Bfr1 in the regulation of P body formation in yeast.
  • To determine how Scp160 and Bfr1 influence the assembly of P body components and their relationship with translation.

Main Methods:

  • Yeast genetics to create deletion mutants (Δscp160, Δbfr1).
  • Immunofluorescence microscopy to visualize P body components like Dcp2.
  • Analysis of polysome profiles to assess translational activity.
  • Treatment with cycloheximide to confirm mRNA presence in observed structures.

Main Results:

  • Loss of Scp160 or Bfr1 leads to the formation of Dcp2-positive foci containing mRNA, independent of stress.
  • Scp160 is essential for the proper assembly of multiple P body components into these foci.
  • In the absence of Bfr1 or Scp160, P body formation is uncoupled from translational attenuation, as polysome profiles remain unchanged.

Conclusions:

  • The proteins Bfr1 and Scp160 function to inhibit P body formation during normal yeast growth.
  • These proteins play a regulatory role in controlling mRNA fate by preventing premature P body assembly.
  • Understanding this mechanism provides insights into the dynamic regulation of mRNA metabolism and cellular homeostasis.
Keywords:
Endoplasmic reticulumProcessing bodiesRibonucleotide particlesSaccharomyces cerevisiaeStress granulesStress responseTranslationmRNAmRNA decaymRNA metabolismmRNA-binding proteins

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