Tertiary structures of mitochondrial tRNAs having characteristic secondary structures.

Some of the animal mitochondrial (mt) tRNAs are thought to have unusual secondary structures as inferred from the mt DNA sequence analysis. To obtain information on the relationship between the unusual secondary and tertiary structures and function of these mt tRNAs at the RNA level, we have analyzed the structure of bovine mt serine tRNAs and obtained the following results. 1) Using the novel secondary structure proposed on the basis of enzymatic probing and phylogenetic comparison and the known tertiary structure of yeast tRNA(Phe), a tertiary structural model of tRNA(Ser) specific for UCU/C/A/G codons was constructed by a computer modeling. In the proposed model, one-base-pair elongation of anticodon stem compensated for the deletions in some of the loop regions of this tRNA, and the model maintained the topological relationship between the anticodon and 3'-CCA terminus as nearly the same as that of yeast tRNA(Phe). The results of chemical modification of this tRNA supported the proposed model. 2) For NMR analysis of tRNA(Ser) specific for AGU/C codons and lacking the D stem, a large scale preparation of the unmodified tRNA transcript has been performed by the use of an in vitro T7 transcription system.