Intramitochondrial sorting of the precursor to yeast cytochrome c oxidase subunit Va.
We have continued our studies on the import pathway of the precursor to yeast cytochrome c oxidase subunit Va (pVa), a mitochondrial inner membrane protein. Previous work on this precursor demonstrated that import of pVa is unusually efficient, and that inner membrane localization is directed by a membrane-spanning domain in the COOH-terminal third of the protein. Here we report the results of studies aimed at analyzing the intramitochondrial sorting of pVa, as well as the role played by ancillary factors in import and localization of the precursor. We found that pVa was efficiently imported and correctly sorted in mitochondria prepared from yeast strains defective in the function of either mitochondrial heat shock protein (hsp)60 or hsp70. Under identical conditions the import and sorting of another mitochondrial protein, the precursor to the beta subunit of the F1 ATPase, was completely defective. Consistent with previous results demonstrating that the subunit Va precursor is loosely folded, we found that pVa could be efficiently imported into mitochondria after translation in wheat germ extracts. This results suggests that normal levels of extramitochondrial hsp70 are also not required for import of the protein. The results of this study enhance our understanding of the mechanism by which pVa is routed to the mitochondrial inner membrane. They suggest that while the NH2 terminus of pVa is exposed to the matrix and processed by the matrix metalloprotease, the protein remains anchored to the inner membrane before being assembled into a functional holoenzyme complex.
Miller
B R
BR
Department of Molecular Biology and Biochemistry, University of California, Irvine 92717.
Cumsky
M G
MG
D002451:Cell Compartmentation; D003576:Electron Transport Complex IV / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000378:metabolism; D046911:Macromolecular Substances; D008928:Mitochondria / Q000201:enzymology* / Q000648:ultrastructure; D012441:Saccharomyces cerevisiae / Q000378:metabolism / Q000648:ultrastructure
D005656:Fungal Proteins; D006360:Heat-Shock Proteins; D046911:Macromolecular Substances; D003576:Electron Transport Complex IV