An exon that prevents transport of a mature mRNA

Proc Natl Acad Sci U S A. 1999 Mar 30;96(7):3813-8. doi: 10.1073/pnas.96.7.3813.

Abstract

In Caenorhabditis elegans, pre-mRNA for the essential splicing factor U2AF65 sometimes is spliced to produce an RNA that includes an extra 216-bp internal exon, exon 3. Inclusion of exon 3 inserts an in-frame stop codon, yet this RNA is not subject to SMG-mediated RNA surveillance. To test whether exon 3 causes RNA to remain nuclear and thereby escape decay, we inserted it into the 3' untranslated region of a gfp reporter gene. Although exon 3 did not affect accumulation or processing of the mRNA, it dramatically suppressed expression of green fluorescent protein (GFP). We showed by in situ hybridization that exon 3-containing gfp RNA is retained in the nucleus. Intriguingly, exon 3 contains 10 matches to the 8-bp 3' splice-site consensus. We hypothesized that U2AF might recognize this octamer and thereby prevent export. This idea is supported by RNA interference experiments in which reduced levels of U2AF resulted in a small burst of gfp expression.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Caenorhabditis elegans / genetics*
  • Caenorhabditis elegans / physiology
  • Exons*
  • Gene Expression Regulation
  • Green Fluorescent Proteins
  • Luminescent Proteins / biosynthesis
  • Luminescent Proteins / genetics
  • Nuclear Proteins*
  • RNA Splicing / genetics*
  • RNA, Messenger / genetics*
  • Recombinant Proteins / biosynthesis
  • Ribonucleoproteins / genetics*
  • Splicing Factor U2AF
  • Transcription, Genetic

Substances

  • Luminescent Proteins
  • Nuclear Proteins
  • RNA, Messenger
  • Recombinant Proteins
  • Ribonucleoproteins
  • Splicing Factor U2AF
  • Green Fluorescent Proteins