Coruzzi G, Tzagoloff A
J Biol Chem. 1979 Sep 25;254(18):9324-30.
A cytoplasmic "petitie" mutant of Saccharomyces cerevisiae (DS200/A1) has been isolated and determined to contain mitochondrial genetic markers in the oxi 1 locus. This locus has previously been reported to code for the structural gene of subunit 2 of cytochrome oxidase (Cabral et al. (1978) J. Biol. Chem. 243, 297-304). The segment of mitochondrial DNA retained in DS200/A1 has a repeat length of approximately 4500 base pairs and based on DNA sequencing contains a 756-nucleotide-long sequence that has been identified as the structural gene of subunit 2 of cytochrome oxidase. The presumptive gene sequence generates an amino acid sequence consistent with the reported molecular weight and composition of subunit 2 of yeast cytochrome oxidase. The correctness of the deduced amino acid sequence is further supported by its extensive homology to the primary structure of bovine cytochrome oxidase. The DNA segment of DS200/A1 has been located on the wild type mitochondrial DNA by comparative restriction mapping. The orientation of the COOH and NH2 termini and the direction of transcription of the gene have been determined.
已分离出酿酒酵母的一种细胞质“小菌落”突变体(DS200/A1),并确定其在oxi 1位点含有线粒体遗传标记。此前有报道称,该位点编码细胞色素氧化酶亚基2的结构基因(卡布拉尔等人,(1978年)《生物化学杂志》243卷,297 - 304页)。保留在DS200/A1中的线粒体DNA片段重复长度约为4500个碱基对,基于DNA测序,其包含一个756个核苷酸长的序列,该序列已被鉴定为细胞色素氧化酶亚基2的结构基因。推测的基因序列产生的氨基酸序列与报道的酵母细胞色素氧化酶亚基2的分子量和组成一致。推导的氨基酸序列的正确性进一步得到其与牛细胞色素氧化酶一级结构广泛同源性的支持。通过比较限制性图谱分析,已确定DS200/A1的DNA片段在野生型线粒体DNA上的位置。还确定了该基因COOH和NH2末端的方向以及转录方向。