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PMID: 2271667 Published · ppublish English Journal Article

The self-splicing RNA of Tetrahymena is trapped in a less active conformation by gel purification.

Biochemistry ·Vol. 29 ·No. 46 ·1990-11-20 ·Pages 10573-6

Walstrum SA, Uhlenbeck OC

Abstract

When the circular form of the self-splicing intervening sequence of Tetrahymena thermophila was purified by denaturing polyacrylamide gel electrophoresis by standard methods, the rate of its reaction with tetrauridylate decreased 150-fold at 30 degrees C and at least 1000-fold at 0 degrees C. The activity of the self-splicing RNA was restored by heating it to high temperature and letting it renature in the presence of Mg2+. The rate of reaction of tetrauridylate with the self-splicing RNA flanked by exons was also greatly decreased by gel purification. The difference in activation energies for the reaction of native and denatured intervening sequences suggests that a substantial conformational rearrangement of the gel-purified RNA occurs prior to reaction.

MeSH Terms
Animals Electrophoresis, Polyacrylamide Gel Kinetics Magnesium/pharmacology Nucleic Acid Conformation Protein Conformation RNA, Catalytic/isolation & purification,metabolism Tetrahymena/genetics,metabolism Thermodynamics
Chemicals
RNA, Catalytic Magnesium
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Walstrum S A
Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309-0215.
Uhlenbeck O C
Article Info
Journal
Biochemistry
Abbr.
Biochemistry
ISSN
0006-2960
Published
1990-11-20
Pages
10573-6
Language
English
Region
United States
NLM ID
0370623
Subset
IM
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