Summary
Detailed mapping localized the PHO 1 mutation between the OLI 2 and OLI 4 loci on mitochondrial DNA of Saccharomyces cerevisiae.
In its mitochondrially integrated form, the PHO 1-ATPase3 was difficult to identify either immunologically or by specific inhibitors like oligomycin and DCCD. Solubilization by Triton X-100 allowed unambiguuous identification of this enzyme as an authentic mitochondrial ATPase. However, Triton extraction produced a 2 to 3 fold enhancement of the PHO 1-ATPase activity which also became drastically cold-sensitive. The wild type ATPase was neither activated nor made cold-labile by solubilization, and retained full sensitivity to oligomycin and DCCD.
Sucrose gradient analysis of the Triton-extracted ATPase from wild type, PHO 1 mutant and rho -strains showed a density difference between the solubilized PHO 1-and wild type ATPase, and similarity between solubilized PHO 1-and rho - ATPase (F1).
Whole cells of the PHO 1 mutant present considerably increased respiration rates.
Comparison of oligomycin-sensitivity in whole cells, coupled isolated mitochondria and membrane-bound ATPase indicates a contrast between oligomycin-resistance of the ATPase and oligomycin-sensitivity of in vivo or in vitro coupling systems, which might characterize the products of this region of mitochondrial DNA.
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Communicated by W. Gajewski
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Somlo, M., Clavilier, L. & Krupa, M. A phantom ATPase in a mitochondrially encoded mutant of Saccharomyces cerevisiae . Molec. Gen. Genet. 156, 289–295 (1977). https://doi.org/10.1007/BF00267184
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DOI: https://doi.org/10.1007/BF00267184