Reconstitution of a group I intron self-splicing reaction with an activator RNA.
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- G van der Horst
- Salk Institute for Biological Studies, La Jolla, CA 92037.
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- A Christian
- Salk Institute for Biological Studies, La Jolla, CA 92037.
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- T Inoue
- Salk Institute for Biological Studies, La Jolla, CA 92037.
Abstract
<jats:p>The self-splicing rRNA intron of Tetrahymena thermophila belongs to a subgroup of group I introns that contain a conserved extra stem-loop structure termed P5abc. A Tetrahymena mutant precursor RNA lacking this P5abc is splicing-defective under standard conditions (5 mM MgCl2/200 mM NH4Cl, pH 7.5) in vitro. However, the mutant precursor RNA by itself is capable of performing the self-splicing reaction without P5abc under different conditions (15 mM MgCl2/2 mM spermidine, pH 7.5). We have investigated the functional role of the P5abc in the mechanism of the self-splicing reaction. When an RNA consisting of the P5abc but lacking the rest of the Tetrahymena intron is incubated with the mutant precursor, the self-splicing reaction proceeds highly efficiently under standard conditions (5 mM MgCl2/200 mM NH4Cl, pH 7.5). Two steps of the bimolecular self-splicing reaction can be performed accurately by a shortened precursor RNA containing all essential components required in the self-splicing reaction and an activator RNA consisting of the P5abc. Gel-mobility-shift assays suggest that two molecules associate by a direct RNA-RNA interaction during the splicing reaction. The results imply that there might exist other small RNAs whose role is to activate ribozymes.</jats:p>
Journal
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- Proceedings of the National Academy of Sciences
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Proceedings of the National Academy of Sciences 88 (1), 184-188, 1991-01
Proceedings of the National Academy of Sciences
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Keywords
Details 詳細情報について
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- CRID
- 1362544418796205440
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- NII Article ID
- 80005648332
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- ISSN
- 10916490
- 00278424
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- Data Source
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- Crossref
- CiNii Articles