Regulated tRNA import in Leishmania mitochondria.

The genes for three new tRNA and a 5S RNA were identified from a genomic DNA clone of 917 nucleotide pairs from the protozoon Leishmania tarentolae. They were encoded in the following order. The transcriptional directions and anticodons are in parentheses: tRNA(Val) (CAC-->)-5SRNA (-->)-tRNA(His) (<--GUG)-tRNA(Phe) (GAA-->). The tRNA(His) and tRNA(Phe) sequences have not been reported previously in trypanosomatid organisms. By northern analysis, tRNA(Val) and tRNA(Phe) were equally distributed between the cytosol and mitochondria, while tRNA(His) was less abundant in mitochondria than in the cytosol. Accordingly, the latter tRNA is classified as Import restricted (Impr). As shown before, 5S RNA was not imported. Recently, Mahapatra and Adhya [S. Mahapatra, T. Ghosh, S. Adhya, Nucl. Acids Res. 22 (1994) 3381-3386; S. Mahapatra, S. Adhya, J. Biol. Chem. 271 (1996) 20432-20437] have developed an in vitro import system in Leishmania and suggested that the D-loop sequence could serve as the import determinant. We examined all available tRNA gene sequences in trypanosomatids but found no apparent consensus within the D-loop that might account for tRNA-import regulation.