Kitakawa M, Grohmann L, Graack H R, Isono K
Department of Biology, Faculty of Science, Kobe University, Japan.
Nucleic Acids Res. 1990 Mar 25;18(6):1521-9. doi: 10.1093/nar/18.6.1521.
The genes for two large subunit proteins, YmL8 and YmL20, of the mitochondrial ribosome of Saccharomyces cerevisiae were cloned by hybridization with synthetic oligonucleotide mixtures corresponding to their N-terminal amino acid sequences. They were termed MRP-L8 and MRP-L20, respectively, and their nucleotide sequences were determined using a DNA sequencer. The MRP-L8 gene was found to encode a 26.8-kDa protein whose deduced amino acid sequence has a high degree of similarity to ribosomal protein L17 of Escherichia coli. The gene MRP-L20 was found to encode a 22.3-kDa protein with a presequence consisting of 18 amino acid residues. By Southern blot hybridization to the yeast chromosomes separated by field-inversion gel electrophoresis, the MRP-L8 and MRP-L20 genes were located on chromosomes X and XI, respectively. Gene disruption experiments indicate that their products, YmL8 and YmL20 proteins, are essential for the mitochondrial function and the absence of these proteins causes instability of the mitochondrial DNA.
通过与对应于酿酒酵母线粒体核糖体两个大亚基蛋白YmL8和YmL20 N端氨基酸序列的合成寡核苷酸混合物杂交，克隆了这两个蛋白的基因。它们分别被命名为MRP-L8和MRP-L20，并使用DNA测序仪测定了它们的核苷酸序列。发现MRP-L8基因编码一种26.8 kDa的蛋白，其推导的氨基酸序列与大肠杆菌核糖体蛋白L17具有高度相似性。发现MRP-L20基因编码一种22.3 kDa的蛋白，其前导序列由18个氨基酸残基组成。通过与场反转凝胶电泳分离的酵母染色体进行Southern印迹杂交，MRP-L8和MRP-L20基因分别位于X染色体和XI染色体上。基因破坏实验表明，它们的产物YmL8和YmL20蛋白对于线粒体功能是必需的，并且这些蛋白的缺失会导致线粒体DNA的不稳定。
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