Complex formation determines the activity of ribozymes directed against potato virus YN genomic RNA sequences
Identifieur interne : 004588 ( Main/Exploration ); précédent : 004587; suivant : 004589Complex formation determines the activity of ribozymes directed against potato virus YN genomic RNA sequences
Auteurs : Renéa. A. Van Der Vlugt [Pays-Bas] ; Marcel Prins [Pays-Bas] ; Rob Goldbach [Pays-Bas]Source :
- Virus Research [ 0168-1702 ] ; 1993.
English descriptors
- Teeft :
- Basepairing, Cleavage, Cleavage activity, Cleavage products, Cleavage reaction, Cleavage reactions, Complementary arms, Complex formation, Detectable cleavage, Efficient cleavage, Equal amounts, Hammerhead, Hammerhead sequence, Hammerhead structure, Hammerhead transcript, Incorrect basepairing, Long arms, Mgcl, Nucleotide, Nucleotide sequence, Plasmid, Potato virus, Reaction samples, Ribozyme, Ribozyme cleavage site, Ribozyme transcript, Ribozymes, Ribozymic activity, Rna, Secondary structure, Secondary structures, Short ribozyme, Substrate rnas, Symons, Transcript, Transcription, Viral, Vlugt.
Abstract
Abstract: A ribozyme was synthesized against a conserved region in the RNA-dependent RNA-polymerase encoding cistron of the important plant pathogen potato virus Y (PVY). This ribozyme was shown to cleave PVY-specific RNA-transcripts efficiently in vitro, with up to 95% of the substrate RNA being cleaved within 2 h incubation at 37°C. A second ribozyme, designed with much shorter viral complementary arms in an attempt to optimize the efficiency of the cleavage reaction, surprisingly failed to cleave the substrates previously cleaved by the longer ribozyme. A much shorter PVY specific RNA-transcript of only 37 nucleotides (nt), however, was cleaved by this short ribozyme proving its ribozymic activity and indicating that the cleavage activity of the ribozyme is, in part, determined by the substrate involved. Analysis of cleavage reactions on non-denaturing polyacrylamide (PAA) gels indicated that incorrect basepairing, interfering with correct formation of the hammerhead structure, was likely to be responsible for the absence of detectable cleavage of the larger substrates by the short ribozyme.
Url:
DOI: 10.1016/0168-1702(93)90081-W
Affiliations:
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A." last="Van Der Vlugt">Renéa. A. 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This ribozyme was shown to cleave PVY-specific RNA-transcripts efficiently in vitro, with up to 95% of the substrate RNA being cleaved within 2 h incubation at 37°C. A second ribozyme, designed with much shorter viral complementary arms in an attempt to optimize the efficiency of the cleavage reaction, surprisingly failed to cleave the substrates previously cleaved by the longer ribozyme. A much shorter PVY specific RNA-transcript of only 37 nucleotides (nt), however, was cleaved by this short ribozyme proving its ribozymic activity and indicating that the cleavage activity of the ribozyme is, in part, determined by the substrate involved. Analysis of cleavage reactions on non-denaturing polyacrylamide (PAA) gels indicated that incorrect basepairing, interfering with correct formation of the hammerhead structure, was likely to be responsible for the absence of detectable cleavage of the larger substrates by the short ribozyme.</div></front></TEI><affiliations><list><country><li>Pays-Bas</li></country><region><li>Gueldre (province)</li></region><settlement><li>Wageningue</li></settlement><orgName><li>Université de Wageningue</li></orgName></list><tree><country name="Pays-Bas"><region name="Gueldre (province)"><name sortKey="Van Der Vlugt, Renea A" sort="Van Der Vlugt, Renea A" uniqKey="Van Der Vlugt R" first="Renéa. A." last="Van Der Vlugt">Renéa. A. Van Der Vlugt</name></region><name sortKey="Goldbach, Rob" sort="Goldbach, Rob" uniqKey="Goldbach R" first="Rob" last="Goldbach">Rob Goldbach</name><name sortKey="Prins, Marcel" sort="Prins, Marcel" uniqKey="Prins M" first="Marcel" last="Prins">Marcel Prins</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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