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Assessment of Selective mRNA Translation in Mammalian Cells by Polysome Profiling
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A programmed -1 ribosomal frameshift signal can function as a cis-acting mRNA destabilizing element

Ewan P Plant1, Pinger Wang, Jonathan L Jacobs

  • 1Department of Cell Biology and Molecular Genetics, Microbiology Building Room 2135, University of Maryland, College Park, MD 20742, USA.

Nucleic Acids Research
|February 6, 2004
PubMed

Insights

Programmed ribosomal frameshifting signals can destabilize mRNA. In yeast, these signals trigger nonsense-mediated mRNA decay (NMD), similar to premature stop codons, offering a new gene regulation mechanism.

Area of Science:

  • Molecular Biology
  • Genetics
  • RNA Biology

Background:

  • Nonsense-mediated mRNA decay (NMD) degrades mRNAs with premature termination codons (PTCs).
  • Programmed -1 ribosomal frameshifting (-1 PRF) is common in viral polycistronic mRNAs but less understood in yeast.
  • Previous studies suggest yeast -1 PRF signals may lead to PTCs.

Purpose of the Study:

  • To investigate if -1 PRF signals act as cis-acting mRNA destabilizing elements in yeast.
  • To determine if -1 PRF targets mRNAs for degradation via the NMD pathway.

Main Methods:

  • Insertion of an L-A viral -1 PRF signal into a PGK1 reporter construct in yeast.
  • Analysis of reporter mRNA half-lives and decay profiles.
  • Correlation analysis between frameshift efficiency and mRNA stability.

Main Results:

  • Even low levels of -1 PRF efficiently targeted reporter mRNA for degradation by NMD.
  • Reporter mRNA half-lives were comparable to those with in-frame PTCs.
  • An inverse correlation was observed between frameshift efficiency and mRNA half-life.
  • NMD activity was confirmed on mRNAs beyond the initial translation round.

Conclusions:

  • -1 PRF signals can function as potent mRNA destabilizing elements in yeast.
  • NMD pathway degrades mRNAs containing -1 PRF signals.
  • Modulating -1 PRF frequency offers a mechanism for post-transcriptional gene regulation.
  • NMD remains active on mRNAs throughout translation in yeast.

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