TARGETING AND TRANSLOCATION OF PROTEINS INTO THE HYDROGENOSOME OF THEPROTIST TRICHOMONAS - SIMILARITIES WITH MITOCHONDRIAL PROTEIN IMPORT

Trichomonads are early-diverging eukaryotes that lack both mitochondri a and peroxisomes. They do contain a double membrane-bound organelle, called the hydrogenosome, that metabolizes pyruvate and produces ATP, To address the origin and biological nature of hydrogenosomes, we have established an in vitro protein import assay. Using purified hydrogen osomes and radiolabeled hydrogenosomal precursor ferredoxin (pFd), we demonstrate that protein import requires intact organelles, ATP and N- ethylmaleimide-sensitive cytosolic factors, Protein import is also aff ected by high concentrations of the protonophore, m-chlorophenylhydraz one (CCCP), Binding and translocation of pFd into hydrogenosomes requi res the presence of an eight amino acid N-terminal presequence that is similar to presequences found an all examined hydrogenosomal proteins . Upon import, pFd is processed to a size consistent with cleavage of the presequence, Mutation of a conserved leucine at position 2 in the presequence to a glycine disrupts import of pFd into the organelle, In terestingly, a comparison of hydrogenosomal and mitochondrial protein presequences reveals striking similarities, These data indicate that m echanisms underlying protein targeting and biogenesis of hydrogenosome s and mitochondria are similar, consistent with the notion that these two organelles arose from a common endosymbiont.