M. Tanudji et al., Signals required for the import and processing of the alternative oxidase into mitochondria, J BIOL CHEM, 274(3), 1999, pp. 1286-1293
The critical residues involved in targeting and processing of the soybean a
lternative oxidase to plant and animal mitochondria was investigated. Impor
t of various site-directed mutants into soybean mitochondria indicated that
positive residues throughout the length of the presequence were important
for import, not just those in the predicted region of amphiphilicity. The p
osition of the positive residues in the C-terminal end of the presequence w
as also important for import. Processing assays of the various constructs w
ith purified spinach mitochondrial processing peptidase showed that all the
-2-position mutants had a drastic effect on processing. In contrast to the
import assay, the position of the positive residue could be changed for pr
ocessing. Deletion mutants confirmed the site-directed mutagenesis data in
that an amphiphilic alpha-helix was not the only determinant of mitochondri
al import in this homologous plant system. Import of these constructs into
rat liver mitochondria indicated that the degree of inhibition differed and
that the predicted region of amphiphilic alpha-helix was more important wi
th rat liver mitochondria. Processing with a rat liver matrix fraction show
ed little inhibition. These results are discussed with respect to targeting
specificity in plant cells and highlight the need to carry out homologous
studies and define the targeting requirements to plant mitochondria.