title: Import of an incompletely folded precursor protein into isolated mitochondria requires an energized inner membrane, but no added ATP.
abstract: We have studied the post-translational import of incomplete precursor chains into isolated yeast mitochondria. The precursor was a fusion protein containing a mitochondrial presequence attached to mouse dihydrofolate reductase. In vitro-synthesis of the precursor was interrupted by the elongation inhibitor cycloheximide and the arrested nascent chains cosedimenting with ribosomes were released by EDTA. These incomplete chains were efficiently imported by isolated yeast mitochondria; their import resembled that of the complete precursor in requiring an energized inner membrane and a mitochondrial presequence. It differed from that of the completed precursor in its resistance to methotrexate (which only binds to correctly folded dihydrofolate reductase) and its independence of added ATP. The incomplete chains were also more sensitive to proteinase K than the completed precursor. We conclude that the incomplete chains were incompletely folded and suggest that the lack of tight folding caused import into mitochondria to become independent of added ATP. This implies that ATP may participate, directly or indirectly, in the unfolding of the precursor for its transport into mitochondria.
authors.affiliation: Biocenter, University of Basel, Switzerland.
authors.forename: K
authors.initials: K
authors.lastname: Verner
authors.forename: G
authors.initials: G
authors.lastname: Schatz
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D003576:Electron Transport Complex IV / Q000235:genetics*; D007425:Intracellular Membranes / Q000378:metabolism*; D051379:Mice; D008928:Mitochondria / Q000378:metabolism*; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D013762:Tetrahydrofolate Dehydrogenase / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D000255:Adenosine Triphosphate; D013762:Tetrahydrofolate Dehydrogenase; D003576:Electron Transport Complex IV