ArticleProceedings of the National Academy of Sciences of the United States of America1999
An important base triple anchors the substrate helix recognition surface within the Tetrahymena ribozyme active site.
Article in Proceedings of the National Academy of Sciences of the United States of America, 1999. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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9 citing papers in PubMed, 36 citations in OpenAlex.
- Sub-3-Å cryo-EM structure of RNA enabled by engineered homomeric self-assembly.Nature methods · 2022Article
- Article
- Structure-function analysis from the outside in: long-range tertiary contacts in RNA exhibit distinct catalytic roles.Biochemistry · 2011Article
- Fluorine substituted adenosines as probes of nucleobase protonation in functional RNAs.Journal of the American Chemical Society · 2008Article
- Comparison of crystal structure interactions and thermodynamics for stabilizing mutations in the Tetrahymena ribozyme.RNA (New York, N.Y.) · 2006Article
- Crystal structure of a group I intron splicing intermediate.RNA (New York, N.Y.) · 2004Article
- Oligonucleotide directed misfolding of RNA inhibits Candida albicans group I intron splicing.Proceedings of the National Academy of Sciences of the United States of America · 2002Article
- Solution structure of an RNA fragment with the P7/P9.0 region and the 3'-terminal guanosine of the tetrahymena group I intron.RNA (New York, N.Y.) · 2002Article
- Detection of N-H...N hydrogen bonding in RNA via scalar couplings in the absence of observable imino proton resonances.Nucleic acids research · 2000Article
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Authors and funding
6 authors at 2 institutions in 2 countries.
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Abstract
Key to understanding the structural biology of catalytic RNA is determining the underlying networks of interactions that stabilize RNA folding, substrate binding, and catalysis. Here we demonstrate the existence and functional importance of a Hoogsteen base triple (U300.A97-U277), which anchors the substrate helix recognition surface within the Tetrahymena group I ribozyme active site. Nucleotide analog interference suppression analysis of the interacting functional groups shows that the U300.A97-U277 triple forms part of a network of hydrogen bonds that connect the P3 helix, the J8/7 strand, and the P1 substrate helix. Product binding and substrate cleavage kinetics experiments performed on mutant ribozymes that lack this base triple (C A-U, U G-C) or replace it with the isomorphous C(+).G-C triple show that the A97 Hoogsteen triple contributes to the stabilization of both substrate helix docking and the conformation of the ribozyme's active site. The U300. A97-U277 base triple is not formed in the recently reported crystallographic model of a portion of the group I intron, despite the presence of J8/7 and P3 in the RNA construct [Golden, B. L., Gooding, A. R., Podell, E. R. & Cech, T. R. (1998) Science 282, 259-264]. This, along with other biochemical evidence, suggests that the active site in the crystallized form of the ribozyme is not fully preorganized and that substantial rearrangement may be required for substrate helix docking and catalysis.
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