Biochemical analysis of pistol self-cleaving ribozymes
| Autor: | Sanshu Li, Kimberly A. Harris, Christina E. Lünse, Kenneth I Brewer, Ronald R. Breaker |
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| Rok vydání: | 2015 |
| Předmět: |
Molecular Sequence Data
Cleavage (embryo) Catalysis Report Consensus Sequence Consensus sequence RNA Catalytic Nucleotide Molecular Biology Ligase ribozyme chemistry.chemical_classification Bacteria Base Sequence biology Nucleotides Ribozyme Computational Biology RNA Kinetics chemistry Biochemistry biology.protein Nucleic Acid Conformation RNA Cleavage Pseudoknot |
| Zdroj: | RNA. 21:1852-1858 |
| ISSN: | 1469-9001 1355-8382 |
| DOI: | 10.1261/rna.052514.115 |
| Popis: | Pistol RNAs are members of a distinct class of self-cleaving ribozymes that was recently discovered by using a bioinformatics search strategy. Several hundred pistol ribozymes share a consensus sequence including 10 highly conserved nucleotides and many other modestly conserved nucleotides associated with specific secondary structure features, including three base-paired stems and a pseudoknot. A representative pistol ribozyme from the bacterium Lysinibacillus sphaericus was found to promote RNA strand scission with a rate constant of ∼10 min−1 under physiological Mg2+ and pH conditions. The reaction proceeds via the nucleophilic attack of a 2′-oxygen atom on the adjacent phosphorus center, and thus adheres to the same general catalytic mechanism of internal phosphoester transfer as found with all other classes of natural self-cleaving ribozymes discovered to date. Analyses of the kinetic characteristics and the metal ion requirements of the cleavage reaction reveal that members of this ribozyme class likely use several catalytic strategies to promote the rapid cleavage of RNA. |
| Databáze: | OpenAIRE |
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