Single VS ribozyme molecules reveal dynamic and hierarchical folding toward catalysis
| Autor: | Shawna L. Hiley, Richard A. Collins, Dominic C. J. Jaikaran, Evgenia N. Nikolova, Nils G. Walter, Miguel J. B. Pereira |
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| Rok vydání: | 2008 |
| Předmět: |
RNA
Untranslated Molecular Sequence Data Biology Catalysis Article Structural Biology Endoribonucleases Fluorescence Resonance Energy Transfer RNA Catalytic Molecular Biology Fluorescent Dyes Base Sequence GIR1 branching ribozyme Ribozyme RNA RNA Fungal Single-molecule FRET Protein tertiary structure GlmS glucosamine-6-phosphate activated ribozyme Neurospora Biochemistry biology.protein Biophysics Mutagenesis Site-Directed Nucleic Acid Conformation Hairpin ribozyme VS ribozyme |
| Zdroj: | Journal of molecular biology. 382(2) |
| ISSN: | 1089-8638 |
| Popis: | Non-coding RNAs of complex tertiary structure are involved in numerous aspects of the replication and processing of genetic information in many organisms; however, an understanding of the complex relationship between their structural dynamics and function is only slowly emerging. The Neurospora Varkud Satellite (VS) ribozyme provides a model system to address this relationship. First, it adopts a tertiary structure assembled from common elements, a kissing loop and two three-way junctions. Second, catalytic activity of the ribozyme is essential for replication of VS RNA in vivo and can be readily assayed in vitro. Here we exploit single molecule FRET to show that the VS ribozyme exhibits previously unobserved dynamic and heterogeneous hierarchical folding into an active structure. Readily reversible kissing loop formation combined with slow cleavage of the upstream substrate helix suggests a model whereby the structural dynamics of the VS ribozyme favor cleavage of the substrate downstream of the ribozyme core instead. This preference is expected to facilitate processing of the multimeric RNA replication intermediate into circular VS RNA, which is the predominant form observed in vivo. |
| Databáze: | OpenAIRE |
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