ISOLATION AND CHARACTERIZATION OF NOVEL OLIGOMYCIN RESISTANT SACCHAROMYCES CEREVISIAE MUTANTS OF THE MITOCHONDRIALLY ENCODED FO ATPASE SUBUNITS
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The mitochondria of Saccharomyces cerevisiae are semi-autonomous organelles that rely on nuclear genes to function. Although most subunits of yeast ATPase are nuclear-encoded, the membrane-bound FO portion is comprised of three mitochondrially-encoded subunits. FO ATPase confers sensitivity to the antibiotic oligomycin, whereby mitochondrial metabolism is disrupted and ATP synthesis ceases. There are four oligomycin resistance loci located on the mitochondrial DNA, namely oli1, oli2, oli3, and oli4. However, there is currently no fine structure genetic map describing these genes. Three hundred and eighty-eight oligomycin-resistant mutants were generated following manganese mutagenesis. Following preliminary testing, mutants were crossed to determine allelism. Random diploid analysis was used to score diploid progeny as either oligomycin-sensitive or oligomycin-resistant, and recombination frequencies were calculated. A fine structure genetic map of the oli1-oli3 region of the mtDNA was generated, and mutants mapping outside this region were identified. Since mitochondria play an essential role in normal cellular growth and respiration, this study and others will help elucidate the resistance genes and the mechanisms of antibiotic binding and organelle function.