Purification and thermal stability of several amino acid-specific tRNAs from an extreme thermophile, Thermus thermophilus HB8.

Three species of methionine tRNAs and phenylalanine, tyrosine, and isoleucine tRNAs were purified from an extreme thermophile, Thermus thermophilus HB8. Formylation studies of the three methionine tRNAs and their codon-specific binding activities to ribosomes showed that two of them (named tRNAf1Met and tRNAf2Met) were initiator tRNAs and the other (named tRNAmMet) was a non-initiator. The tRNAs from T. thermophilus all had melting temperatures of up to ten degrees higher than the corresponding species from E. coli. Most of the species also had slightly higher G+C contents than the corresponding species of E. coli, and each of them contained one mol each of the modified nucleosides, O2'-methylguanosine (Gm), 2-thioribothymidine (s2T), and 1-methyladenosine (m1A). Their high melting temperatures could be explained by their high G+C contents and the presence of the modified nucleosides, espically s2T. Comparison of the melting temperatures of T. thermophilus tRNAf2Met with those of E. coli tRNAfMet and tRNAmMet at different magnesium concentrations showed that magnesium was also a factor in the thermostability of the thermophile tRNA.