title: The first twelve amino acids (less than half of the pre-sequence) of an imported mitochondrial protein can direct mouse cytosolic dihydrofolate reductase into the yeast mitochondrial matrix.
abstract: Yeast cytochrome c oxidase subunit IV (an imported mitochondrial protein) is made as a larger precursor with a transient pre-sequence of 25 amino acids. If this pre-sequence is fused to the amino terminus of mouse dihydrofolate reductase (a cytosolic protein) the resulting fusion protein is imported into the matrix space, and cleaved to a smaller size, by isolated yeast mitochondria. We have now fused progressively shorter amino-terminal segments of the subunit IV pre-sequence to dihydrofolate reductase and tested each fusion protein for import into the matrix space and cleavage by the matrix-located processing protease. The first 12 amino acids of the subunit IV pre-sequence were sufficient to direct dihydrofolate reductase into the mitochondrial matrix, both in vitro and in vivo. However, import of the corresponding fusion protein into the matrix was no longer accompanied by proteolytic processing. Fusion proteins containing fewer than nine amino-terminal residues from the subunit IV pre-piece were not imported into isolated mitochondria. The information for transporting attached mouse dihydrofolate reductase into mitochondria is thus contained within the first 12 amino acids of the subunit IV pre-sequence.
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D003600:Cytosol / Q000201:enzymology; D003576:Electron Transport Complex IV / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005796:Genes; D005800:Genes, Fungal; D046911:Macromolecular Substances; D051379:Mice; D008928:Mitochondria / Q000201:enzymology*; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics*
chemical_list: D046911:Macromolecular Substances; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D013762:Tetrahydrofolate Dehydrogenase; D003576:Electron Transport Complex IV