Boutry M, Vassarotti A, Ghislain M, Douglas M, Goffeau A
J Biol Chem. 1984 Mar 10;259(5):2840-4.
The structural genes for the two major subunits of the mitochondrial ATPase were isolated among genomic clones from the yeast Schizosaccharomyces pombe by transformation and complementation of mutants unable to grow on glycerol and lacking either the alpha or the beta subunits. The plasmid pMa1 containing a 2.3-kilobase genomic insert transformed the mutant A23-13 lacking a detectable alpha subunit. The transformant grew on glycerol and contained an alpha subunit of normal electrophoretic mobility. The plasmid pMa2 containing a 5.4-kilobase genomic insert transformed the mutant B59-1 lacking the beta subunit. The transformant grew on glycerol and contained a beta subunit of normal mobility. The structural gene for the beta ATPase subunit for the fission yeast S. pombe was localized within the pMa2 insert by hybridization to a probe containing the beta ATPase gene from the budding yeast Saccharomyces cerevisiae (Saltzgaber, J., Kunapuli, S., and Douglas, M. G. (1983) J. Biol. Chem. 258, 11465-11470). The mRNAs which hybridized to pMa1 and pMa2 were translated by a reticulocyte lysate into polypeptides of Mr = 59,000 and 54,000, respectively. These genes products reacted with an anti-F1-ATPase serum and therefore correspond most probably to precursors of the alpha and beta subunits.
通过对不能在甘油上生长且缺乏α或β亚基的突变体进行转化和互补，从粟酒裂殖酵母的基因组克隆中分离出线粒体ATP酶两个主要亚基的结构基因。含有2.3千碱基基因组插入片段的质粒pMa1转化了缺乏可检测到的α亚基的突变体A23 - 13。转化体在甘油上生长，并含有电泳迁移率正常的α亚基。含有5.4千碱基基因组插入片段的质粒pMa2转化了缺乏β亚基的突变体B59 - 1。转化体在甘油上生长，并含有迁移率正常的β亚基。通过与含有来自酿酒酵母的β - ATP酶基因的探针杂交，将粟酒裂殖酵母β - ATP酶亚基的结构基因定位在pMa2插入片段内（萨尔茨加伯，J.，库纳普利，S.，和道格拉斯（1983年）《生物化学杂志》258，11465 - 11470）。与pMa1和pMa2杂交的mRNA在网织红细胞裂解物中分别被翻译成分子量为59,000和54,000的多肽。这些基因产物与抗F1 - ATP酶血清发生反应，因此很可能对应于α和β亚基的前体。
