Primary structure of bovine liver tRNATrp

Identity elements in bovine tRNATrp required for the specific stimulation of gelonin, a plant ribosome-inactivating protein

“…Trp and U-G in yeast tRNA Trp ) and the triple interaction of nt 9 with bp 23-12 (G-C-G and G-A-U, respectively) differ in the two tRNAs+ Table 1 reports previous data (Brigotti et al+, 1996), largely confirmed in our laboratory, for the geloninstimulating activity of native mammalian and yeast tRNA Trp + The twofold difference in activity between beef and rabbit tRNA Trp was explained by differences in the methods of preparation and storage of the two tRNAs obtained in two different laboratories+ Sequencing data had, in fact, long ago established the similarity of the two tRNAs (Fournier et al+, 1978) and the conservation of tRNA Trp in avian and mammalian cells has since been well documented (Sprinzl et al+, 1996)+ As shown in Table 1, the lack of nucleotide modifications in the synthetic transcript of bovine tRNA Trp induced no loss of gelonin-stimulating activity, which actually rose to that previously reported for rabbit tRNA+ In contrast, both the native and the synthetic yeast tRNA…”

“…tRNA genes containing the T7-promoter directly adjacent to guanosine ϩ1 of the tRNA were constructed using six overlapping DNA oligonucleotides (Sampson & Uhlenbeck, 1988)+ The oligonucleotides were purchased from Genenco (Florence, Italy) and purified by electrophoresis on denaturing 20% polyacrylamide gels followed by gel filtration through P2 columns (1 ϫ 18 cm) equilibrated and eluted with water+ The oligonucleotides used to construct the wild-type bovine and yeast tRNA genes are shown in Figure 4+ The primary structure of bovine tRNA Trp chosen for the synthetic transcript was that reported by Sprinzl et al+ (1996) and contains, at the heterogeneity points of native tRNA (Fournier et al+, 1978;Keith & Heyman, 1990), A rather than G at position 2, U rather than C at positions 16 and 47, G rather than A at position 57, and A rather than G at position 59+ Yeast tRNA Trp has no such points of heterogeneity (Keith et al+, 1972)+ Assembly of oligonucleotides by phosphorylation (20 mL separate reaction mixtures), annealing, and ligation (100 mL reaction mixture) was performed as described by Sampson and Uhlenbeck (1988)…”

Mutual conformational changes of tryptophanyl‐tRNA synthetase and tRNATrp in the course of their specific interaction

“…Moreover, this approach has been used with a homologous beef tryptophan pair and with a heterologous pair (yeast tRNATrp and beef enzyme). We have chosen these tRNAsTrp since they have similar parameters in the aminoacylation reaction catalyzed by beef tryptophanyltRNA synthetase [15] although their primary structures are quite different [16,171.…”

Role of exposed cytosine residues in aminoacylation activity of tRNATrp

“…Fig.1. The primary structure of tRNATrp (beef) [21]. Symbols designate cytidine residues that are converted into uridine by modification with bisultite under the conditions when the 3-dimensional structure of the tRNA is preserved [9].…”