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Science 7 February 1997:
Vol. 275. no. 5301, pp. 847 - 849
DOI: 10.1126/science.275.5301.847
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Reports

Joining the Two Domains of a Group I Ribozyme to Form the Catalytic Core

Michael A. Tanner and Thomas R. Cech *

Self-splicing group I introns, like other large catalytic RNAs, contain structural domains. Although the crystal structure of one of these domains has been determined by x-ray analysis, its connection to the other major domain that contains the guanosine-binding site has not been known. Site-directed mutagenesis and kinetic analysis of RNA splicing were used to identify a base triple in the conserved core of both a cyanobacterial (Anabaena) and a eukaryotic (Tetrahymena) group I intron. This long-range interaction connects a sequence adjacent to the guanosine-binding site with the domain implicated in coordinating the 5' splice site helix, and it thereby contributes to formation of the active site. The resulting five-strand junction, in which a short helix forms base triples with three separate strands in the Tetrahymena intron, reveals exceptionally dense packing of RNA.

Department of Chemistry and Biochemistry, Howard Hughes Medical Institute, University of Colorado, Boulder, Colorado 80309-0215, USA.
*   To whom correspondence should be addressed.

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THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Crystal structure of a group I intron splicing intermediate.
P. L. ADAMS, M. R. STAHLEY, M. L. GILL, A. B. KOSEK, J. WANG, and S. A. STROBEL (2004)
RNA 10, 1867-1887
   Abstract »    Full Text »    PDF »
A base triple in the Tetrahymena group I core affects the reaction equilibrium via a threshold effect.
K. KARBSTEIN, K.-H. TANG, and D. HERSCHLAG (2004)
RNA 10, 1730-1739
   Abstract »    Full Text »    PDF »
Distinct sites of phosphorothioate substitution interfere with folding and splicing of the Anabaena group I intron.
A. Luptak and J. A. Doudna (2004)
Nucleic Acids Res. 32, 2272-2280
   Abstract »    Full Text »    PDF »
Structure-function relationships of two closely related group IC3 intron ribozymes from Azoarcus and Synechococcus pre-tRNA.
Y. Ikawa, D. Naito, H. Shiraishi, and T. Inoue (2000)
Nucleic Acids Res. 28, 3269-3277
   Abstract »    Full Text »    PDF »
An important base triple anchors the substrate helix recognition surface within the Tetrahymena ribozyme active site.
A. A. Szewczak, L. Ortoleva-Donnelly, M. V. Zivarts, A. K. Oyelere, A. V. Kazantsev, and S. A. Strobel (1999)
PNAS 96, 11183-11188
   Abstract »    Full Text »    PDF »
A Preorganized Active Site in the Crystal Structure of the Tetrahymena Ribozyme.
B. L. Golden, A. R. Gooding, E. R. Podell, and T. R. Cech (1998)
Science 282, 259-264
   Abstract »    Full Text »
The Therapeutic Potential of Ribozymes.
H. A. James and I. Gibson (1998)
Blood 91, 371-382
   Abstract »    Full Text »    PDF »
Defining the chemical groups essential for Tetrahymena group I intron function by nucleotide analog interference mapping.
S. A. Strobel and K. Shetty (1997)
PNAS 94, 2903-2908
   Abstract »    Full Text »    PDF »
Site-directed Mutagenesis of the Yeast Resolving Enzyme Cce1 Reveals Catalytic Residues and Relationship with the Intron-splicing Factor Mrs1.
B. N. Wardleworth, M. Kvaratskhelia, and M. F. White (2000)
J. Biol. Chem. 275, 23725-23728
   Abstract »    Full Text »    PDF »


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Science. ISSN 0036-8075 (print), 1095-9203 (online)