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Ectopic 5' splice sites inhibit gene expression by engaging RNA surveillance and silencing pathways in plants

Krzysztof Wypijewski1, Csaba Hornyik, Jane A Shaw

  • 1Plant Pathology Department, Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, United Kingdom.

Plant Physiology
|August 12, 2009
PubMed

Insights

Altering mRNA processing in plants by adding splice sites triggers pre-mRNA degradation and gene silencing. This RNA quality control mechanism, involving short-interfering RNAs (siRNAs), is conserved across kingdoms.

Area of Science:

  • Molecular Biology
  • Plant Science
  • RNA Biology

Background:

  • mRNA maturation involves quality control to degrade aberrant pre-mRNAs.
  • Viruses utilize host RNA quality control for gene regulation.
  • Conserved mechanisms between kingdoms are crucial for understanding fundamental biology.

Purpose of the Study:

  • To investigate the impact of 5' splice sites in the 3'-untranslated region on pre-mRNA processing in plants.
  • To explore the role of RNA quality control and silencing pathways in response to aberrant pre-mRNAs.
  • To elucidate the cross-talk between RNA quality control, 3'-end maturation, and RNA silencing.

Main Methods:

  • Reporter gene assays in plants.
  • Analysis of pre-mRNA cleavage and polyadenylation.
  • Quantification of short-interfering RNAs (siRNAs) using sequencing.
  • Gene expression analysis in wild-type and mutant backgrounds (RDR6-deficient).
  • Coexpression of silencing suppressors.

Main Results:

  • Inclusion of 5' splice sites in the 3'-untranslated region led to pre-mRNA degradation and inhibition of homologous gene expression.
  • Aberrant pre-mRNA processing preferentially accumulated 24-nucleotide (nt) siRNAs, suggesting involvement of the silencing machinery.
  • Gene expression was not restored in RDR6-deficient plants or with silencing suppressor coexpression, despite reduced 24-nt siRNA accumulation.

Conclusions:

  • Plant RNA quality control machinery can degrade aberrant pre-mRNAs, a mechanism conserved across kingdoms.
  • A complex interplay exists between RNA quality control, 3'-end pre-mRNA maturation, and RNA silencing pathways.
  • These pathways can discriminate between different types of aberrant RNAs, influencing gene expression regulation.

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