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Competition for XPO5 binding between Dicer mRNA, pre-miRNA and viral RNA regulates human Dicer levels

Yamina Bennasser1, Christine Chable-Bessia, Robinson Triboulet

  • 1Centre National de la Recherche Scientifique, Institut de Génétique Humaine UPR1142, Montpellier, France.

Nature Structural & Molecular Biology
|February 8, 2011
PubMed

Insights

Exportin-5 inhibition downregulates Dicer, a key enzyme for microRNA (miRNA) maturation. This cross-regulation impacts cellular physiology and viral replication by affecting Dicer mRNA nuclear export.

Area of Science:

  • Molecular Biology
  • Gene Regulation
  • RNA Interference

Background:

  • MicroRNAs (miRNAs) are crucial regulators of gene expression.
  • The RNA interference (RNAi) pathway involves multiple protein and RNA components.
  • Exportin-5 mediates the nuclear export of pre-miRNAs, essential for miRNA biogenesis.

Purpose of the Study:

  • To investigate the cross-regulatory relationship between exportin-5 and Dicer within the RNAi machinery.
  • To elucidate the molecular mechanism underlying the interaction between exportin-5 and Dicer mRNA.

Main Methods:

  • Inhibition of exportin-5 in human cells.
  • Analysis of Dicer mRNA and protein levels.
  • In vitro binding assays and cellular RNA immunoprecipitation (RIP) to assess RNA-protein interactions.
  • Overexpression studies to titrate exportin-5 levels.
  • Adenoviral infection models.

Main Results:

  • Exportin-5 inhibition led to decreased Dicer expression via post-transcriptional mechanisms.
  • Dicer mRNA exhibited increased nuclear localization upon exportin-5 inhibition.
  • Exportin-5 was shown to directly bind to Dicer mRNA.
  • Overexpression of pre-miRNA or viral RNA saturated exportin-5, disrupting its interaction with Dicer mRNA and reducing Dicer levels.
  • Adenoviral infection mimicked this saturation, enhancing viral replication.

Conclusions:

  • A novel cross-regulatory mechanism exists between pre-miRNAs/viral RNAs and Dicer mediated by exportin-5.
  • Exportin-5 plays a critical role in regulating Dicer expression by controlling Dicer mRNA nuclear export.
  • This interaction is crucial for maintaining cellular homeostasis and can be exploited by viruses for replication.

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