CHEMICAL AMINOACYLATION OF TRANSFER-RNAS

  • 1 January 1978
    • journal article
    • research article
    • Vol. 253  (13) , 4517-4520
Abstract
Incubation of abbreviated tRNA (tRNA-C-COH) with (chemically) preaminoacylated P1,P2-di(adenosine 5''-)diphosphates in the presence of purified [Escherichia coli] RNA ligase effected transfer of an aminoacyladenylate moiety to the 3''-terminus of the abbreviated tRNA in good yield. Aminoacylated (or misacylated) tRNA may thus be prepared from fractionated or unfractionated tRNA-C-COH; each of the 5 aminoacylated dinucleoside diphosphates tested was utilized as a substrate by RNA ligase. That the resulting chemically aminoacylated tRNA were identical with those prepared by enzymatic aminoacylation was judged by comparison of chromatographic properties on benzolated DEAE-cellulose, rates of chemical deacylation and affinities for elongation factor Tu, and the ability of misacylated tRNA so derived to be deacylated chemically and then reactivated enzymatically with their cognate amino acids.