Miyakawa I, Okazaki-Higashi C, Higashi T, Furutani Y, Sando N
Department of Biology, Faculty of Science, Yamaguchi University, Japan.
Plant Cell Physiol. 1996 Sep;37(6):816-24. doi: 10.1093/oxfordjournals.pcp.a029017.
Mitochondrial (mt) nucleoids were isolated with a high degree of purity from the yeast Pichia jadinii, in which the mitochondrial DNA (mtDNA) is linear. Field-inversion gel electrophoresis (FIGE) revealed that significant amounts of mtDNA could be isolated intact, as linear molecules of 41 kbp, from the isolated mt-nucleoids. Fifteen different proteins were detected in the mt-nucleoid fraction and, eight of these proteins bound to DNA. The patterns of mt-nucleoid proteins and of the DNA-binding proteins after gel electrophoresis in the presence of SDS were somewhat different from those of such proteins from Saccharomyces cerevisiae. The corresponding proteins isolated from the mt-nucleoids of four other species of yeast in the genera Pichia and Williopsis also differed from one another in terms of electrophoretic mobility in the presence of SDS. In immunoblotting experiments, antibodies that had been raised against the 67-kDa protein of mt-nucleoids from S.cerevisiae and the YMN-1 monoclonal antibody that is specific for a 48-kDa protein in the mt-nucleoids from S. cerevisiae did not recognize any proteins in the mt-nucleoids from Pichia jadinii and four other species of yeast. The results suggest the considerable diversity of the proteins in the mt-nucleoids of yeasts.
从线粒体DNA（mtDNA）呈线性的酵母季也蒙毕赤酵母中，以高纯度分离出了线粒体（mt）类核。场反转凝胶电泳（FIGE）显示，从分离出的mt类核中，可以完整地分离出大量作为41 kbp线性分子的mtDNA。在mt类核组分中检测到了15种不同的蛋白质，其中8种蛋白质与DNA结合。在SDS存在下进行凝胶电泳后，mt类核蛋白和DNA结合蛋白的模式与酿酒酵母中的此类蛋白略有不同。从毕赤酵母属和威克酵母属的其他四种酵母的mt类核中分离出的相应蛋白质，在SDS存在下的电泳迁移率方面也彼此不同。在免疫印迹实验中，针对酿酒酵母mt类核的67 kDa蛋白产生的抗体以及对酿酒酵母mt类核中48 kDa蛋白具有特异性的YMN-1单克隆抗体，均未识别季也蒙毕赤酵母和其他四种酵母的mt类核中的任何蛋白质。结果表明酵母mt类核中的蛋白质具有相当大的多样性。
