Self-splicing of a group I intron reveals partitioning of native and misfolded RNA populations in yeast

Scott A. Jackson, Sujatha Koduvayur, Sarah A. Woodson

Research output: Contribution to journalArticle

Abstract

Stable RNAs must form specific three-dimensional structures, yet many RNAs become kinetically trapped in misfolded conformations. To understand the factors that control the accuracy of RNA folding in the cell, the self-splicing activity of the Tetrahymena group I intron was compared in different genetic contexts in budding yeast. The extent of splicing was 98% when the intron was placed in its natural rDNA context, but only 3% when the intron was expressed in an exogenous pre-mRNA. Further experiments showed that the probability of forming the active intron structure depends on local sequence context and transcription by Pol I. Pre-rRNAs decayed at similar rates, whether the intron was wild type or inactivated by an internal deletion, suggesting that most of the unreacted pre-rRNA is incompetent to splice. Northern blots and complementation assays showed that mutations that destabilize the intron tertiary structure inhibited self-splicing and processing of internal transcribed spacer 2. The data are consistent with partitioning of pre-rRNAs into active and inactive populations. The misfolded RNAs are sequestered and degraded without refolding to a significant extent. Thus, the initial fidelity of folding can dictate the intracellular fate of transcripts containing this group I intron. Published by Cold Spring Harbor Laboratory Press.

Original languageEnglish (US)
Pages (from-to)2149-2159
Number of pages11
JournalRNA
Volume12
Issue number12
DOIs
StatePublished - Dec 1 2006

Keywords

  • Metastable structure
  • Pre-rRNA processing
  • RNA folding
  • RNA stability

ASJC Scopus subject areas

  • Molecular Biology

Fingerprint Dive into the research topics of 'Self-splicing of a group I intron reveals partitioning of native and misfolded RNA populations in yeast'. Together they form a unique fingerprint.

  • Cite this