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PMID: 2643083 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Effects of substrate structure on the kinetics of circle opening reactions of the self-splicing intervening sequence from Tetrahymena thermophila: evidence for substrate and Mg2+ binding interactions.

Nucleic acids research ·Vol. 17 ·No. 1 ·1989-01-11 ·Pages 355-71

Sugimoto N, Tomka M, Kierzek R, Bevilacqua PC, Turner DH

Abstract

The self-splicing intervening sequence from the precursor rRNA of Tetrahymena thermophila cyclizes to form a covalently closed circle. This circle can be reopened by reaction with oligonucleotides or water. The kinetics of circle opening as a function of substrate and Mg2+ concentrations have been measured for dCrU, rCdU, dCdT, and H2O addition. Comparisons with previous results for rCrU suggest: (1) the 2' OH of the 5' sugar of a dinucleoside phosphate is involved in substrate binding, and (2) the 2' OH of the 3' sugar of a dimer substrate is involved in Mg2+ binding. Evidently, the binding site for a required Mg2+ ion is dependent on both the ribozyme and the dimer substrate. The apparent activation energy and entropy for circle opening by hydrolysis are 31 kcal/mol and 50 eu, respectively. The large, positive activation entropy suggests a partial unfolding of the ribozyme is required for reaction.

MeSH Terms
Animals Dinucleoside Phosphates/metabolism Introns Kinetics Magnesium/metabolism Mathematics Models, Theoretical RNA Precursors/genetics,metabolism RNA Splicing RNA, Catalytic RNA, Ribosomal/genetics,metabolism Tetrahymena/genetics Thermodynamics
Chemicals
Dinucleoside Phosphates RNA Precursors RNA, Catalytic RNA, Ribosomal Magnesium
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Sugimoto N
Department of Chemistry, University of Rochester, NY 14627.
Tomka M
Kierzek R
Bevilacqua P C
Turner D H
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/17.1.355
Published
1989-01-11
Pages
355-71
Language
English
Region
England
NLM ID
0411011
PMCID
PMC331555
Subset
IM
Grants
NIGMS NIH HHS · GM22989 · United States
External Links
PubMed source DOI: 10.1093/nar/17.1.355 PMC Full Text View journal details
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