Nucleic Acids Research, Vol 26, Issue 9 2037-2041, Copyright © 1998 by Oxford University Press
S Mahapatra, S Ghosh, SK Bera, T Ghosh, A Das and S Adhya
Transfer RNAs are selectively imported from the cytoplasm into mitochondria
of kinetoplastid protozoa such as Leishmania . The specific structural
features of tRNA which determine selectivity are largely unknown. Using an
in organello system from Leishmania , the import signals on tRNATyrand on a
synthetic transcript which binds to the same receptor, were studied by
deletion and reconstruction analyses. In both cases, short
oligoribonucleotides (minihelices) containing the sequence UGGYAGAG were
imported with high efficiency in the presence of ATP. This motif is present
in the D arm of tRNATyr, as well as in the majority of imported Leishmania
tRNAs. Deletion of the D arm, or a point mutation in the conserved motif,
reduces importability. The import signal coincides with the binding site
for the mitochondrial receptor TAB. tRNAGln, which is not imported, forms
non-productive, TAB- independent complexes with the mitochondrial surface.
However, the observation that the imported:bound ratio of the D arm
minihelix is higher than that of the entire molecule suggests that the
post-binding translocation step is constrained in terms of size or
structural flexibility. Kinetic studies of minihelix import indicate
stepwise insertion of the molecule into import channels.
ARTICLES
The D arm of tRNATyr is necessary and sufficient for import into Leishmania mitochondria in vitro
Genetic Engineering Laboratory, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Calcutta 700032, India.
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