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Título: | Effects of the trinucleotide preceding the self-cleavage site on eggplant latent viroid hammerheads: differences in co- and post-transcriptional self-cleavage may explain the lack of trinucleotide AUC in most natural hammerheads |
Autor: | Carbonell, Alberto CSIC ORCID ; Peña, Marcos de la ; Flores, Ricardo CSIC ORCID; Gago, Selma CSIC ORCID | Fecha de publicación: | 16-sep-2006 | Editor: | Oxford University Press | Citación: | Nucleic Acids Research 34/19:5613–5622 (2006) | Resumen: | Eggplant latent viroid (ELVd) can form stable hammerhead structures in its (1) and ( ) strands. These ribozymes have the longest helices I reported in natural hammerheads, with that of the ELVd (1) hammerhead being particularly stable (5/7 bp are G-C). Moreover, the trinucleotide preceding the self-cleavage site of this hammerhead is AUA, which together with GUA also found in some natural hammerheads, deviate from the GUC present in most natural hammerheads including the ELVd ( ) hammerhead. When the AUA trinucleotide preceding the self-cleavage site of the ELVd (1) hammerhead was substituted by GUA and GUC, as well as by AUC (essentially absent in natural hammerheads), the values of the self-cleavage rate constants at low magnesium of the purified hammerheads were: ELVd-(1)-AUC ELVd-(1)-GUC.ELVd-(1)-GUA. ELVd-(1)-AUA. However, the ELVd-(1)-AUC hammerhead was the catalytically less efficient during in vitro transcription, most likely because of the transient adoption of catalytically-inactive metastable structures. These results suggest that natural hammerheads have been evolutionary selected to function co-transcriptionally, and provide a model explaining the lack of trinucleotide AUC preceding the self-cleavage site of most natural hammerheads. Comparisons with other natural hammerheads showed that the ELVd-(1)-GUC and ELVd-(1)-AUC hammerheads are the catalytically most active in a post-transcriptional context with low magnesium | URI: | http://hdl.handle.net/10261/49157 | DOI: | 10.1093/nar/gkl717 | E-ISSN: | 1362-4962 |
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