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 lnteractions

Naoki Sugimoto, Mary Tomka, Ryszard Kicrzek, Philip C. Bevilacqua, Douglas H. Turner

Research output: Contribution to journalArticle

57 Citations (Scopus)

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 Mg 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.

Original languageEnglish (US)
Pages (from-to)355-371
Number of pages17
JournalNucleic acids research
Volume17
Issue number1
DOIs
StatePublished - Jan 11 1989

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Tetrahymena thermophila
Catalytic RNA
Entropy
Introns
Dinucleoside Phosphates
RNA Precursors
Oligonucleotides
Hydrolysis
Binding Sites
Ions
Water

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

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title = "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 lnteractions",
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 Mg 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.",
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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 lnteractions. / Sugimoto, Naoki; Tomka, Mary; Kicrzek, Ryszard; Bevilacqua, Philip C.; Turner, Douglas H.

In: Nucleic acids research, Vol. 17, No. 1, 11.01.1989, p. 355-371.

Research output: Contribution to journalArticle

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AB - 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 Mg 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.

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