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Related Experiment Videos

3' poly(A) is dispensable for translation.

A M Searfoss1, R B Wickner

  • 1Laboratory of Biochemistry and Genetics, National Institute of Diabetes, Digestive and Kidney Diseases, National Institutes of Health, 8 Center Drive, MSC 0830, Bethesda, MD 20892-0830, USA.

Proceedings of the National Academy of Sciences of the United States of America
|August 2, 2000
PubMed
Summary

The poly(A) tail is crucial for mRNA translation and stability. However, nonpoly(A) mRNA can be efficiently translated, a process normally repressed by Ski2p and Slh1p proteins.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Virology

Background:

  • The 3' poly(A) tail is essential for mRNA translation and stability in wild-type cells.
  • Mutations in the superkiller (ski) genes enhance the expression of polyadenylation-negative (poly(A)(-)) mRNAs from yeast RNA viruses.
  • Ski2p, a DEVH-box RNA helicase, and Slh1p, a protein with similar functions, both repress the copy number of L-A double-stranded RNA (dsRNA) viruses.

Purpose of the Study:

  • To investigate the functional overlap between Ski2p and Slh1p.
  • To determine if nonpoly(A) mRNA can be translated in the absence of these proteins.
  • To elucidate the role of Ski2p and Slh1p in regulating mRNA translation and stability.

Main Methods:

  • Creation and analysis of slh1Delta ski2Delta double mutant yeast strains.

Related Experiment Videos

  • Assessment of cellular mRNA turnover rates in mutant strains.
  • Electroporation of nonpoly(A) mRNA into yeast strains and analysis of translation kinetics.

    • Main Results:

    • slh1Delta ski2Delta double mutants are viable and exhibit normal cellular mRNA turnover.
    • These double mutants efficiently translate electroporated nonpoly(A) mRNA with kinetics similar to polyadenylation-positive (poly(A)(+)) mRNA.
    • The translation machinery is capable of utilizing nonpoly(A) mRNA when the repressive action of Ski2p and Slh1p is removed.

    Conclusions:

    • Ski2p and Slh1p are nonessential proteins that normally repress the translation of nonpoly(A) mRNA.
    • The yeast translation apparatus can efficiently use nonpoly(A) mRNA, but this capability is masked by the action of Ski2p and Slh1p.
    • These findings reveal a regulatory mechanism controlling mRNA translation independent of the poly(A) tail.