Rescue of an abasic hairpin ribozyme by cationic nucleobases: evidence for a novel mechanism of RNA catalysis.
Chem Biol
; 9(4): 465-73, 2002 Apr.
Article
em En
| MEDLINE
| ID: mdl-11983335
ABSTRACT
The hairpin ribozyme catalyzes a reversible phosphodiester cleavage reaction. We examined the roles of conserved nucleobases in catalysis using an abasic ribozyme rescue strategy. Loss of the active site G8 nucleobase reduced the cleavage rate constant by 350-fold while loss of A9 and A10 nucleobases reduced activity less than 10-fold. Certain heterocyclic amines restored partial activity when provided in solution to the variant lacking G8. Heterocyclic amines that were capable of rescue shared the exocyclic amine and cyclic amide in common with the Watson-Crick hydrogen bonding face of guanine. In contrast to the shallow pH dependence of unmodified ribozyme activity, rescue activity increased sharply with decreasing pH. These results support a novel model for RNA catalysis in which a cationic nucleobase contributes electrostatic stabilization to negative charge developing in the transition state.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Sequência de Bases
/
RNA Catalítico
Idioma:
En
Revista:
Chem Biol
Ano de publicação:
2002
Tipo de documento:
Article