Pol II-directed short RNAs suppress the nuclear export of mRNA

Plant Mol Biol. 2010 Dec;74(6):591-603. doi: 10.1007/s11103-010-9700-x. Epub 2010 Oct 17.

Abstract

The synthesis and subsequent nuclear export of non-coding RNA (ncRNA) directed by RNA polymerase (Pol) II is very sensitive to abiotic and biotic external stimuli including pathogen challenges. To assess whether stress-induced ncRNAs may suppress the nuclear export of mRNA, we exploited the ability of Agrobacterium tumefaciens to co-deliver Pol I, II and III promoter-based vectors for the transcription of short (s) ncRNAs, GFP mRNA or genomic RNA of plant viruses (Tobacco mosaic virus, TMV; or Potato virus X, PVX) into the nucleus of Nicotiana benthamiana cells. We showed that, in contrast to Pol I- and Pol III-derived sncRNAs, all tested Pol II-derived sncRNAs (U6 RNA, tRNA or artificial RNAs) resulted in decreased expression of GFP and host mRNA. The level of this inhibitory effect depended on the non-coding transcript length and promoter strength. Short coding RNA (scRNA) can also compete with mRNA for nuclear export. We showed that scRNA, an artificial 117-nt short sequence encoding Elastin-Like peptide element tandems with FLAG sequence (ELF) and the 318-nt N. benthamiana antimicrobial peptide thionin (defensin) gene efficiently decreased GFP expression. The stress-induced export of Pol II-derived sncRNA and scRNA into the cytoplasm via the mRNA export pathway may block nucleocytoplasmic traffic including the export of mRNA responsible for antivirus protection. Consistent with this model, we observed that Pol II-derived sncRNAs as well as scRNA, thionin and ELF strongly enhanced the cytoplasmic reproduction of TMV and PVX RNA.

Publication types

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

MeSH terms

  • Active Transport, Cell Nucleus
  • Biological Transport
  • Cell Nucleus / metabolism*
  • Cytoplasm / metabolism
  • Green Fluorescent Proteins / analysis
  • Nicotiana / genetics*
  • Potexvirus / genetics
  • RNA Polymerase II / physiology*
  • RNA, Messenger / metabolism*
  • RNA, Small Interfering / analysis
  • RNA, Small Interfering / metabolism*
  • RNA, Small Untranslated / analysis
  • RNA, Small Untranslated / physiology
  • RNA, Viral / metabolism
  • Tobacco Mosaic Virus / genetics

Substances

  • RNA, Messenger
  • RNA, Small Interfering
  • RNA, Small Untranslated
  • RNA, Viral
  • Green Fluorescent Proteins
  • RNA Polymerase II