The synthesis and functional evaluation of RNA and DNA polymers having the sequence of Escherichia coli tRNA(fMet).

European Journal of Biochemistry
J P PerreaultR Cedergren

Abstract

Stepwise, solid-phase chemical synthesis has provided long RNA and DNA polymers related to the sequence of Escherichia coli tRNA(fMet). The 34-ribonucleotide oligomer corresponding to the sequence of the 5'-half tRNA molecule has been synthesized and then characterized by gel purification, terminal nucleotide determinations and sequence analysis. This 34-nucleotide oligomer serves as an acceptor in the RNA-ligase-catalyzed reaction with a phosphorylated 43-ribonucleotide oligomer corresponding to the sequence of the 3'-half molecule of tRNA(fMet). The DNA molecule having the sequence of tRNA(fMet) is a 76-deoxyribonucleotide oligomer with a 3'-terminal riboadenosine residue and all U residues replaced by T. These polymers have been compared with an oligodeoxyribonucleotide lacking all 2'-hydroxyl groups except for the 3'-terminal 2'-OH, an oligoribonucleotide lacking modified nucleosides and E. coli tRNA(fMet). The all-RNA 77-nucleotide oligomer can be aminoacylated by E. coli methionyl-tRNA synthetase preparation from E. coli with methionine and threonylated in the A37 position using a yeast extract. In agreement with work by Khan and Roe using tDNA(Phe) and tDNA(Lys), the rA77-DNA(fMet) can be aminoacylated, and preliminary e...Continue Reading

References

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Citations

Mar 29, 2000·Biopolymers·P J Beuning, K Musier-Forsyth
May 1, 1997·Nature Biotechnology·R R Breaker
Apr 30, 1990·European Journal of Biochemistry·J PaquetteR Cedergren
Oct 1, 1995·Chemistry & Biology·R R Breaker, G F Joyce
Nov 1, 1994·Bioconjugate Chemistry·S M Hecht
Dec 26, 2001·The Journal of Biological Chemistry·Susanne T NonekowskiGeorge A Garcia
Jun 15, 1995·Nature·B Cuenoud, J W Szostak
Jan 29, 2000·Journal of Natural Products·S M Hecht

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