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

Plant Physiol. 2009 Oct;151(2):955-65. doi: 10.1104/pp.109.139733. Epub 2009 Aug 7.

Abstract

The quality control of mRNA maturation is a highly regulated process that surveys pre-mRNA integrity and eliminates improperly matured pre-mRNAs. In nature, certain viruses regulate the expression of their genes by hijacking the endogenous RNA quality control machinery. We demonstrate that the inclusion of 5' splice sites within the 3'-untranslated region of a reporter gene in plants alters the pre-mRNA cleavage and polyadenylation process, resulting in pre-mRNA degradation, exemplifying a regulatory mechanism conserved between kingdoms. Altered pre-mRNA processing was associated with an inhibition of homologous gene expression in trans and the preferential accumulation of 24-nucleotide (nt) short-interfering RNAs (siRNAs) as opposed to 21-nt siRNA subspecies, suggesting that degradation of the aberrant pre-mRNA involves the silencing machinery. However, gene expression was not restored by coexpression of a silencing suppressor or in an RNA-dependent RNA polymerase (RDR6)-deficient background despite reduced 24-nt siRNA accumulation. Our data highlight a complex cross talk between the quality control RNA machinery, 3'-end pre-mRNA maturation, and RNA-silencing pathways capable of discriminating among different types of aberrant RNAs.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions / genetics
  • Gene Expression Regulation, Plant*
  • Gene Knockdown Techniques
  • Genes, Plant
  • Green Fluorescent Proteins / metabolism
  • Mutagenesis, Insertional
  • Nicotiana / genetics*
  • Nucleic Acid Conformation
  • Polyadenylation
  • RNA Interference*
  • RNA Splice Sites / genetics*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA, Plant / metabolism*
  • RNA, Small Interfering / chemistry
  • RNA, Small Interfering / metabolism
  • Suppression, Genetic
  • Tombusvirus / genetics

Substances

  • 3' Untranslated Regions
  • RNA Splice Sites
  • RNA, Messenger
  • RNA, Plant
  • RNA, Small Interfering
  • Green Fluorescent Proteins