核背景和体内环境在人成纤维细胞mtDNA复制关键控制位点衰老依赖性T414G突变的可变分离行为中的作用

Y Michikawa, K Laderman, K Richter, G Attardi
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引用次数: 13

摘要

先前的研究表明,在大多数65岁以上的人类受试者中,在mtDNA复制的关键控制位点,非遗传性T414G翻转大量积累(高达50%的mtDNA),而在年轻人中则没有。在本研究中,对几个携带异质T414G突变的成纤维细胞群体进行长期体外培养,发现突变细胞由野生型细胞生长。这一观察结果支持了之前的结论,即突变积累是一种体内现象,同时也表明突变型细胞与野生型细胞之间存在内在的生理差异。此外,亚克隆实验揭示了该突变在原始成纤维细胞群体中惊人的马赛克分布,表明其以异质或同质形式存在于一小部分(18-32%)成纤维细胞中,而在其他成纤维细胞中不存在。在其他研究中,线粒体从携带突变的成纤维细胞转移到mtdna缺失的143B。然而,TK- rho0206细胞显示了突变的马赛克分布的持久性,几乎完全转变为同质性。后一种现象的普遍性将排除转化实验中一个或几个线粒体的奠基者效应,而更倾向于指出核背景在T414G突变的体外行为中的重要作用。rho0细胞转化体中同质突变的稳定性为分析其生化效应提供了有力的工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Role of nuclear background and in vivo environment in variable segregation behavior of the aging-dependent T414G mutation at critical control site for human fibroblast mtDNA replication.

Previous work had shown a large accumulation (up to 50% of mtDNA) of a noninherited T414G transversion at a critical control site for mtDNA replication in skin fibroblasts from the majority of human subjects above 65 years old, and its absence in younger individuals. In the present studies, long-term in vitro culture of several fibroblasts populations carrying the heteroplasmic T414G mutation revealed an outgrowth of the mutant cells by wild-type cells. This observation supported the previous conclusion that the mutation accumulation is an in vivo phenomenon, while, at the same time, indicating intrinsic physiological differences between mutant and wild-type cells. Furthermore, subcloning experiments revealed a striking mosaic distribution of the mutation in the original fibroblasts populations, as shown by its presence, in heteroplasmic or homoplasmic form, in a fraction (18-32%) of the fibroblasts, and its absence in the others. In other investigations, transfer of mitochondria from mutation-carrying fibroblasts into mtDNA-less 143B.TK- rho0 206 cells revealed the persistence of the mosaic distribution of the mutation, however, with a near-complete shift to homoplasmy. The generality of the latter phenomenon would exclude a founder effect by one or few mitochondria in the transformation experiments, and would rather point to the important role of the nuclear background in the in vitro behavior of the T414G mutation. The stability of the homoplasmic mutation in rho0 cell transformants provides a powerful tool for analyzing its biochemical effects.

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