Dynamics of ribozyme binding of substrate revealed by fluorescence-detected stopped-flow methods

Philip C. Bevilacqua, Ryszard Kierzek, Kenneth A. Johnson, Douglas H. Turner

Research output: Contribution to journalArticle

176 Scopus citations

Abstract

Fluorescence-detected stopped-flow and equilibrium methods have been used to study the mechanism for binding of pyrene (pyr)-labeled RNA oligomer substrates to the ribozyme (catalytic RNA) from Tetrahymena thermophila. The fluorescence of these substrates increases up to 25-fold on binding to the ribozyme. Stopped-flow experiments provide evidence that pyr experiences at least three different microenvironments during the binding process. A minimal mechanism is presented in which substrate initially base pairs to ribozyme and subsequently forms tertiary contacts in an RNA folding step. All four microscopic rate constants are measured for ribozyme binding of pyrCCUCU.

Original languageEnglish (US)
Pages (from-to)1355-1358
Number of pages4
JournalScience
Volume258
Issue number5086
DOIs
StatePublished - 1992

All Science Journal Classification (ASJC) codes

  • General

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