Flexibility of the Bulge Formed Between a Hairpin Ribozyme and Deoxy-Substrate Analogues
Identifieur interne : 004665 ( Main/Exploration ); précédent : 004664; suivant : 004666Flexibility of the Bulge Formed Between a Hairpin Ribozyme and Deoxy-Substrate Analogues
Auteurs : D. V. Dossantos [France] ; J. L. Fourrey [France] ; A. Favre [France]Source :
- Biochemical and Biophysical Research Communications [ 0006-291X ] ; 1993.
Abstract
Abstract: The conformation of the bulge formed between the hairpin ribozyme R derived from (−)sTRSV and noncleavable all-deoxy-substrate analogues dS was studied by photoaffinity labelling. The photolabel deoxy-6-thioinosine was inserted in place of residue G+1 or A−1, located immediately 3′ and 5′ to the cleavage site, respectively. Upon 335 nm irradiation both substrate analogues were linked to ribozyme at multiple sites. Formation of the R-dS complex is absolutely required for the generation of the crosslinks, since they were detected neither in the absence of Mg2+ nor upon using a ds6l containing 14-mer, unable to interact with the ribozyme. The fraction of ribozyme crosslinked at completion of the reaction increased with increasing analogue concentrations, yielding apparent KD values for the R-dS complex in the range of 5 ± 2 μM. Multiple crosslinks between ribozyme and each one of the substrate analogues provide clear evidence for a large flexibility of the bulge region.
Url:
DOI: 10.1006/bbrc.1993.1058
Affiliations:
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The photolabel deoxy-6-thioinosine was inserted in place of residue G+1 or A−1, located immediately 3′ and 5′ to the cleavage site, respectively. Upon 335 nm irradiation both substrate analogues were linked to ribozyme at multiple sites. Formation of the R-dS complex is absolutely required for the generation of the crosslinks, since they were detected neither in the absence of Mg2+ nor upon using a ds6l containing 14-mer, unable to interact with the ribozyme. The fraction of ribozyme crosslinked at completion of the reaction increased with increasing analogue concentrations, yielding apparent KD values for the R-dS complex in the range of 5 ± 2 μM. Multiple crosslinks between ribozyme and each one of the substrate analogues provide clear evidence for a large flexibility of the bulge region.</div></front></TEI><affiliations><list><country><li>France</li></country><region><li>Île-de-France</li></region><settlement><li>Gif-sur-Yvette</li></settlement></list><tree><country name="France"><region name="Île-de-France"><name sortKey="Dossantos, D V" sort="Dossantos, D V" uniqKey="Dossantos D" first="D. V." last="Dossantos">D. V. Dossantos</name></region><name sortKey="Favre, A" sort="Favre, A" uniqKey="Favre A" first="A." last="Favre">A. Favre</name><name sortKey="Fourrey, J L" sort="Fourrey, J L" uniqKey="Fourrey J" first="J. L." last="Fourrey">J. L. Fourrey</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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