The isolation from ribonucleic acid of substituted uridines containing alpha-aminobutyrate moieties derived from methionine

Abstract

The RNA of an established line of Chinese hamster cells growing in cell culture contains a small number of uridines substituted at the 3-position with a gamma-linked alpha-aminobutyrate residue. Structure has been ascertained by: (a) examination of incorporation of isotopic labels from precursors; (b) degradation with anhydrous hydrazine and comparison of the products with synthetic material or with hydrazinolysis products of known uridines; and (c) comparison of the unknowns as their hydantoin derivatives with the 5-beta-(bromoethyl)hydantoin alkylation products of uridine and of 1- and 3-methylpseudouridine. In this manner it is shown that 18-S RNA of ribosomes contains a single residue of a nucleoside which we tentatively identify as 1-methyl-3-gamma-(alpha-amino-alpha-carboxypropyl)pseudouridine per molecule. RNA isolated from the supernatant fraction, sedimenting at 4 S and co-electrophoresing with transfer RNA on polyacrylamide gels, contains several similar bases, one of which is identified as 3-gamma-(alpha-amino-alpha-carboxypropyl)uridine. Each of the above nucleosides derives its alpha-aminobutyrate residue from methionine.