The structure and aminoacylation of a temperature-sensitive tRNATrp (Escherichia coli).
A temperature-sensitive (t.s.) tRNATrp from Escherichia coli has a single base change from the wild type (w.t.) species, which results in the loss of a base pair at the bottom of the CCA stem of the cloverleaf structure. Thermodynamic studies on this t.s. tRNA show that it is more susceptible to denaturation than the w.t. due to a larger change in the entropy of denaturation. Correlated with this thermodynamic result is the finding that the denatured t.s. tRNA's T psi C loop is more susceptible to digestion by T1 RNase, suggesting that it has greater freedom than the corresponding structure on the denatured w.t. molecule. In contrast, the native form of the t.s. tRNATrp is very similar to the w.t. with regard to aminoacylation, T1 RNase susceptibility, and column chromatographic mobility, despite the fact that it necessarily has one less base pair. In addition, the well known denaturation-dependent shift in column chromatographic mobility, which is observed for both the t.s. and w.t. molecules, depends on a modification in the anticodon loop, since tRNATrp lacking that modification does not shift when denatured. Thus, though it is not usually thought to be implicated, denaturation probably affects the conformation of the anticodon loop. The lethal phenotype of the mutant at high temperature, defective attenuation of the tryptophan biosynthetic operon in the mutant, and some aspects of the denatured state are clarified by these findings.
D000215:Acylation; D001483:Base Sequence; D004926:Escherichia coli / Q000378:metabolism*; D007700:Kinetics; D008274:Magnesium; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D009695:Nucleic Acid Renaturation; D009843:Oligoribonucleotides / Q000032:analysis; D012343:RNA, Transfer / Q000378:metabolism; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism; D006163:Ribonuclease T1; D013816:Thermodynamics; D014364:Tryptophan
D009843:Oligoribonucleotides; D012346:RNA, Transfer, Amino Acyl; D014364:Tryptophan; D012343:RNA, Transfer; D006163:Ribonuclease T1; D008274:Magnesium