Fluorescence of covalently attached pyrene as a general RNA folding probe

Mary K. Smalley, Scott K. Silverman

Research output: Contribution to journalArticle

Abstract

Fluorescence techniques are commonly and powerfully applied to monitor biomolecular folding. In a limited fashion, the fluorescence emission intensity of covalently attached pyrene has been used as a reporter of RNA conformational changes. Here, we pursue two goals: we examine the relationship between tether identity and fluorescence response, and we determine the general utility of pyrene fluorescence to monitor RNA folding. The P4-P6 domain of the Tetrahymena group I intron RNA was systematically modified at multiple nucleotide positions with pyrene derivatives that provide a range of tether lengths and compositions between the RNA and chromophore. Certain tethers typically lead to a superior fluorescence signal upon RNA folding, as demonstrated by equilibrium titrations with Mg2+. In addition, useful fluorescence responses were obtained with pyrene placed at several nucleotide positions dispersed throughout P4-P6. This suggests that monitoring of tertiary folding by fluorescence of covalently attached pyrene will be generally applicable to structured RNA molecules.

Original languageEnglish (US)
Pages (from-to)152-166
Number of pages15
JournalNucleic acids research
Volume34
Issue number1
DOIs
StatePublished - Jan 1 2006

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ASJC Scopus subject areas

  • Genetics

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