Abundance of tRNA^Phe lacking the peroxy Y-base in mouse neuroblastoma

Abstract

Affinity chromatography on anti-Y (Y is a tricyclic imidazopurine to which is attached a complex four-carbon side chain) antibody immobilized to Sepharose was used to determine the proportion of rat liver tRNAPhe species containing the peroxy Y-nucleoside. Unfractionated Unfractionated mammalian tRNA was aminoacylated with labeled phenylalanine. The phenylalanyl-tRNA was then chemically acetylated to yield N-acetylphenylalanyl-tRNA. When this preparation was applied to the antibody column, between 6-10% of the radioactivity was not bound to the column, indicating a deficiency of peroxy Y-nuceloside in a minor isoaccepting tRNAPhe species. In contrast to normal tissues (including embryonic tissue), about 85% of the tRNAPhe from mouse neuroblastoma C-1300 or N-18 tumors lack the peroxy Y-base, a property which is not affected by tumor age. Rat liver labeled N-acetylphenylalanyl-tRNA preparations were resolved on Plaskon chromatography (RPC-5) into two minor peaks closely followed by a mojor component. A high proportion of the two minor tRNAPhe species was unable to bind to anti-Y antibodies. Upon mild acid treatment, the minor and major tRNAPhe species eluted simultaneously from Plaskon columns, at a much reduced salt concentration. These results would indicate that the two minor tRNAPhe species from rat liver as well as the major component contain a tricyclic imidazopurine base that differs from each other in its side chain. About 85% of the N-acetylphenylalanyl-tRNA from neuroblastoma was resolved by Plaskon chromatography as an early eluting peak. The position of this major neuroblastoma tRNAPhe species was not altered by mild acid treatment, and its elution position from the column almost coincides with that of acid-treated normal rat liver tRNAPhe. The latter results would suggest that most of the tRNAPhe chains from neuroblastoma lack the tricyclic imidazopurine of normal rat liver tRNAPhe, but are very close if not identical in primary nucleotide sequence.

Comparison of rat liver and Walker 256 carcinosarcoma tRNAs

“…The earlier work in this area has been reviewed [7] and more recent reports of alterations in. tRNA chromatographic mobility have been published [5,[8][9][10][11][12][13][14][15][16][17][18][19][20][21][22]. Only in a few instances [5,[8][9][10][11][12][13] has the specific alteration, been investigated fully enough to identify the structural change which caused the observed chromatographic shift in.…”

Role of Shwachman-Bodian-Diamond syndrome protein in translation machinery and cell chemotaxis: a comparative genomics approach

“…The UUU phenylalanine codon is highly prone to frameshift in the 3′ (rightward) direction at pyrimidine 3′ contexts,19 and wybutosine supports reading frame maintenance by stabilizing codon–anticodon interactions during decoding on the ribosome 20,21. Wybutosine synthesis might proceed through sequential reactions in a multiple protein complex assembled with the precursor tRNA and may be linked to exosomal/proteosomal structure that includes the SBDS protein.…”

Functionally impaired tRNA from ethionine treated rats as detected in injected Xenopus oocytes

“…The tRNA was extracted with two volumes of water-saturated phenol and acid-insoluble radioactivity was determined as described above. Plaskon chromatography Plaskon (RPC 5) chromatography was carried out as previously described (16,17) with 200 ml linear gradients of: 0.6M-0.67M NaCl for the separation of N-acetylphenylalanyl-tRNA, 0.45M-0.55M NaCl for the separation of N-acetylhistidyl-tRNA and 0.4M-0.6M NaCl for N-acetylmethionyl-tRNA. All the NaCl solutions contained 0.01M NaOAc (pH 4.5) and 0.01M MgCl2.…”

“…It was previously demonstrated that Adomet is involved in the biosynthesis of the hypermodified Wye and Q bases in tRNAPhe and tRNAHis respectively (23,24,25). The proportion of these modified bases varies between tRNA from normal and several tumor lines and can be analyzed by Plaskon chromatography (16,17,(24)(25)(26)(27)(28)(29)(30). One way of explaining the ethionine effect is to assume that it affects the biosynthesis of these bases.…”

“…The percentage of tRNAPhe deficient in peroxy-Wye base was also determined by affinity chromatography on an anti-Wye antibody column (14,17) (Fig.3). The two tRNA preparations showed an identical profile on the antibody column confirming the results obtained by Plaskon chromatography.…”