Inhibition of methylated nucleoside synthesis in vivo: accumulation of incompletely methylated transfer RNA in ethionine-treated cells of Escherichia coli B

Abstract

tRNA prepared from cells of E. coli B that had been incubated with 0.5% DL-ethionine (Ethio sRNA) was found to accept methyl groups from 14CH3-S-adenosyl-methionine in the enzymatic reaction catalyzed in vitro by tRNA methyl transferases from untreated cells of the same organism. tRNA from cells that were not exposed to ethionine did not accept a significant level of methyl groups when incubated with the same enzyme system. Base ratio analysis of the product obtained after in vitro addition of methyl groups to Ethio sRNA by enzymes from normal E. coli B indicated that a high proportion of uracil sites in this tRNA were available for enzymatic methylation. These results indicated that tRNA from ethionine-treated organisms was recognized by the homologous enzymes to be incompletely methylated, while, as previously shown, all methyl-acceptor sites on tRNA from normal cells were already filled, and that Ethio sRNA was preferentially deficient in methyl groups on uracil moieties in the RNA molecules. Ethionine thus appears to interfere with normal tRNA modification in vivo.