Structural comparison of human, bovine, rat, and Walker 256 carcinosarcoma asparaginyl-tRNA

Abstract

The complete nucleotide sequences of human placenta, human liver, and bovine liver tRNA Asn have been determined. A comparison of these tRNA structures with the previously reported nucleotide sequences of rat liver and Walker 256 carcinosarcoma tRNA Asn reveals that the primary nucleotide sequences of the major species of mammalian cytoplasmic tRNA Asn are conserved in higher eucaryotes. The complete nucleotide sequence of these tRNAs is: pG-U-C-U-C-U-G-U-m 1G-m 2G-C-G-C-A-A-D-C-G-G-D-X-A-G-C-G-C-m 2 2G- ψ-ψ-C-G-G-C-U-Q(G)-U-U-t 6A-A-C-C-G-A-A-A-G-m 7G-D-U-G-G-U-G-G-Z- ψ-C-G-m 1A-G-C-C-C-A-C-C-C-A-G-G-G-A-C-G-C-C-A OH where X is 3-(3- amino-3- carboxyl-n- propyl) uridine , Q is 7-(4,5- cis-dihydroxyl-1-cyclopenten-3-yl-aminomethyl)-7-deazaguanosine, Z is an unknown modified nucleotide, and Q(G) represents the replacement of Q nucleoside by G nucleoside in Walker 256 carcinosarcoma tRNA Asn. These primary structures were determined by combined use of the 3H- and 32P-post-labeling techniques. Sequences were compared by tritium nucleoside trialcohol analysis, complete RNAase T 1 digestion followed by 3H-labeled fingerprinting on polyethylenimine-impregnated cellulose by two-dimensional thin-layer chromatography (TLC), and polyacrylamide gel electrophoresis of either 5′- 32P- and/or 3′-[ 32P]pCp-labeled tRNA after partial ribonuclease digestions.