Chemical synthesis of a biologically active natural tRNA with its minor bases.
AUTOR(ES)
Gasparutto, D
RESUMO
The complete chemical synthesis of an E. coli tRNA(Ala) with its specific minor nucleosides, dihydrouridine, ribothymidine and pseudouridine, is reported. The method makes use of protected 2'-O-tertiobutyldimethylsilyl-ribonucleoside-3'-O-(2-cyanoethyl-N- ethyl-N- methyl)phosphoramidites. The exocyclic amino functions of the bases were protected by the phenoxyacetyl group for purines and acetyl for cytosine. The assembling has been performed on a silica support with coupling yield better than 98% within 2 min of condensation. Triethylamine tris-hydrofluoride allowed a clean and complete deprotection of the tBDMS groups. The synthetic tRNA(Ala) has been transcribed into cDNA by reverse transcriptase and sequenced. With E. coli alanyl-tRNA synthetase the alanyl acceptance activity and kcat/Km were 672 pmol/A260 and 6 x 10(4)M-1s-1, respectively.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=334300Documentos Relacionados
- 1H NMR of valine tRNA modified bases. Evidence for multiple conformations.
- A potential primer for reverse transcription of mdg3, a Drosophila copia-like element, is a leucine tRNA lacking its 3' terminal 5 bases.
- A biologically active 53 kDa fragment of overproduced alanyl-tRNA synthetase from Thermus thermophilus HB8 specifically interacts with tRNA Ala acceptor helix.
- Synthesis and NMR of RNA with selective isotopic enrichment in the bases.
- Biologically active fragment of a human tRNA synthetase inhibits fluid shear stress-activated responses of endothelial cells
