Fast folding of a ribozyme by stabilizing core interactions: Evidence for multiple folding pathways in RNA

Jie Pan, Michael L. Deras, Sarah A. Woodson

Research output: Contribution to journalArticle

Abstract

Folding of the Tetrahymena ribozyme under physiological conditions in vitro is limited by slow conversion of long-lived intermediates to the active structure. These intermediates arise because the most stable domain of the ribozyme folds 10-50 times more rapidly than the core region containing helix P3. Native gel electrophoresis and time-resolved X-ray-dependent hydroxyl radical cleavage revealed that mutations that weaken peripheral interactions between domains accelerated folding five-fold, while a point mutation that stabilizes P3 enabled 80% of the mutant RNA to reach the native conformation within 30 seconds at 22°C. The P3 mutation increased the folding rate of the catalytic core as much as 50-fold, so that both domains of the ribozyme were formed at approximately the same rate. The results show that the ribozyme folds rapidly without significantly populating metastable intermediates when native interactions in the ribozyme core are stabilized relative to peripheral structural elements. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)133-144
Number of pages12
JournalJournal of Molecular Biology
Volume296
Issue number1
DOIs
StatePublished - Feb 11 2000
Externally publishedYes

Fingerprint

Catalytic RNA
RNA
Tetrahymena
Mutation
Point Mutation
Hydroxyl Radical
Electrophoresis
Catalytic Domain
Gels
X-Rays

Keywords

  • Group I intron
  • Hydroxyl radical cleavage
  • RNA folding
  • X-ray footprinting

ASJC Scopus subject areas

  • Virology

Cite this

Fast folding of a ribozyme by stabilizing core interactions : Evidence for multiple folding pathways in RNA. / Pan, Jie; Deras, Michael L.; Woodson, Sarah A.

In: Journal of Molecular Biology, Vol. 296, No. 1, 11.02.2000, p. 133-144.

Research output: Contribution to journalArticle

Pan, Jie ; Deras, Michael L. ; Woodson, Sarah A. / Fast folding of a ribozyme by stabilizing core interactions : Evidence for multiple folding pathways in RNA. In: Journal of Molecular Biology. 2000 ; Vol. 296, No. 1. pp. 133-144.
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