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Measurement of mRNA Decay Rates in Saccharomyces cerevisiae Using rpb1-1 Strains
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Global analysis of mRNA decay intermediates in Saccharomyces cerevisiae

Yuriko Harigaya1, Roy Parker

  • 1Howard Hughes Medical Institute, Department of Molecular and Cellular Biology, University of Arizona, Tucson, AZ 85721, USA. Yuriko.Harigaya@Colorado.EDU

Proceedings of the National Academy of Sciences of the United States of America
|July 4, 2012
PubMed

Insights

Endonucleases play a minor role in yeast mRNA decay, with Dcp2 being the primary decapping enzyme. Researchers also found a novel mRNA discard pathway during pre-mRNA splicing.

Area of Science:

  • Molecular Biology
  • Yeast Genetics
  • RNA Metabolism

Background:

  • Eukaryotic mRNA decay typically involves deadenylation and 3' to 5' degradation or decapping.
  • Endonucleolytic cleavage sites have been identified in metazoan mRNAs, but their role in yeast is less understood.

Purpose of the Study:

  • To investigate the role of endonucleases in mRNA degradation in Saccharomyces cerevisiae.
  • To identify alternative mRNA decay pathways and regulatory mechanisms.

Main Methods:

  • Mapping 5' monophosphate ends on mRNAs in wild-type and dcp2 xrn1 yeast cells.
  • Stabilizing mRNA endonuclease cleavage products to facilitate detection.
  • Analyzing pre-mRNA splicing intermediates.

Main Results:

  • Endonucleolytic cleavage was observed at low levels on only a few mRNAs, indicating a minor role in yeast mRNA decay.
  • An unknown mechanism, independent of known decapping enzymes, generates 5' exposed ends on some mRNAs, though Dcp2 is the primary decapping enzyme.
  • Debranched lariat intermediates from intron-containing genes were identified, revealing a significant mRNA discard pathway during pre-mRNA splicing.

Conclusions:

  • Endonucleases are not a major factor in Saccharomyces cerevisiae mRNA decay.
  • A novel pathway for mRNA degradation or regulation exists, potentially linked to pre-mRNA splicing.
  • Dcp2 is the principal enzyme responsible for mRNA decapping in yeast.

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