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hUPF2 silencing identifies physiologic substrates of mammalian nonsense-mediated mRNA decay

Jürgen Wittmann1, Elly M Hol, Hans-Martin Jäck

  • 1Division of Molecular Immunology, Department of Internal Medicine III, Nikolaus Fiebiger Center, University of Erlangen-Nürnberg, Glückstrasse 6, D-91054 Erlangen, Germany.

Molecular and Cellular Biology
|February 2, 2006
PubMed

Insights

Nonsense-mediated mRNA decay (NMD) degrades faulty mRNAs. Silencing UPF2 revealed NMD also regulates normal gene expression by controlling specific physiologic transcripts, impacting cellular dynamics.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Nonsense-mediated mRNA decay (NMD) is a crucial eukaryotic surveillance pathway.
  • NMD primarily degrades aberrant mRNAs containing premature termination codons (PTCs).
  • The role of NMD in regulating physiologic transcripts in mammals remains incompletely understood.

Purpose of the Study:

  • To identify novel physiologic NMD substrates.
  • To investigate the impact of NMD silencing on cellular dynamics.
  • To elucidate the function of hUPF2 in NMD regulation.

Main Methods:

  • Stable down-regulation of hUPF2 using RNA interference in HeLa cells.
  • Assessment of NMD-mediated degradation of aberrant PTC transcripts.
  • Analysis of hUPF1 phosphorylation.
  • Genome-wide DNA microarray expression profiling.

Main Results:

  • hUPF2 silencing impaired recognition of aberrant PTC transcripts.
  • hUPF2 silencing did not affect cell growth or viability.
  • hUPF2 silencing diminished hUPF1 phosphorylation, suggesting a regulatory role.
  • 37 novel transcripts were up-regulated and 57 down-regulated in hUPF2-silenced cells.
  • Approximately 60% of up-regulated mRNAs possessed typical NMD motifs.

Conclusions:

  • NMD is essential for maintaining transcriptome integrity by eliminating aberrant transcripts.
  • NMD plays a significant role in the posttranscriptional control of specific physiologic transcripts.
  • hUPF2 modulates NMD activity, potentially through regulation of hUPF1 phosphorylation.

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