Effect of transcription on folding of the Tetrahymena ribozyme

Susan L. Heilman-Miller, Sarah A. Woodson

Research output: Contribution to journalArticle

Abstract

Sequential formation of RNA interactions during transcription can bias the folding pathway and ultimately determine the functional state of a transcript. The kinetics of cotranscriptional folding of the Tetrahymena L-21 ribozyme was compared with refolding of full-length transcripts under the same conditions. Sequential folding after transcription by phage T7 or Escherichia coli polymerase is only twice as fast as refolding, and the yield of native RNA is the same. By contrast, a greater fraction of circularly permuted variants folded correctly at early times during transcription than during refolding. Hybridization of complementary oligonucleotides suggests that cotranscriptional folding enables a permuted RNA beginning at G303 to escape nonnative interactions in P3 and P9. We propose that base pairing of upstream sequences during transcription elongation favors branched secondary structures that increase the probability of forming the native ribozyme structure.

Original languageEnglish (US)
Pages (from-to)722-733
Number of pages12
JournalRNA
Volume9
Issue number6
DOIs
StatePublished - Jun 1 2003

Keywords

  • E. coli RNA polymerase
  • Group I intron
  • In vitro transcription
  • Native gel electrophoresis
  • RNA structure
  • Sequential folding
  • T7 RNA polymerase

ASJC Scopus subject areas

  • Molecular Biology

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  • Cite this

    Heilman-Miller, S. L., & Woodson, S. A. (2003). Effect of transcription on folding of the Tetrahymena ribozyme. RNA, 9(6), 722-733. https://doi.org/10.1261/rna.5200903