Targeting and translocation of proteins into the hydrogenosome of the protist Trichomonas: similarities with mitochondrial protein import.
Trichomonads are early-diverging eukaryotes that lack both mitochondria 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 hydrogenosomes 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 affected by high concentrations of the protonophore, m-chlorophenylhydrazone (CCCP). Binding and translocation of pFd into hydrogenosomes requires the presence of an eight amino acid N-terminal presequence that is similar to presequences found on 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. Interestingly, a comparison of hydrogenosomal and mitochondrial protein presequences reveals striking similarities. These data indicate that mechanisms underlying protein targeting and biogenesis of hydrogenosomes and mitochondria are similar, consistent with the notion that these two organelles arose from a common endosymbiont.
Bradley
P J
PJ
Department of Microbiology & Immunology, School of Medicine, Molecular Biology Institute, University of California, Los Angeles 90095, USA.
Lahti
C J
CJ
Plümper
E
E
Johnson
P J
PJ
D000255:Adenosine Triphosphate / Q000378:metabolism; D000477:Alkylating Agents / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology; D005288:Ferredoxins / Q000378:metabolism; D007476:Ionophores / Q000494:pharmacology; D007930:Leucine; D008928:Mitochondria / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D011498:Protein Precursors / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D015800:Protozoan Proteins / Q000378:metabolism*; D013696:Temperature; D014244:Trichomonas / Q000187:drug effects / Q000378:metabolism*
D000477:Alkylating Agents; D005288:Ferredoxins; D007476:Ionophores; D011498:Protein Precursors; D015800:Protozoan Proteins; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D000255:Adenosine Triphosphate; D007930:Leucine; D005033:Ethylmaleimide