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Science 20 November 1992:
Vol. 258. no. 5086, pp. 1355 - 1358
DOI: 10.1126/science.1455230
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Articles

Science, Vol 258, Issue 5086, 1355-1358
Copyright © 1992 by American Association for the Advancement of Science

articles

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

PC Bevilacqua, R Kierzek, KA Johnson, and DH Turner

Department of Chemistry, University of Rochester, NY 14627.

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.


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