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酿酒酵母中DNA从线粒体逃逸至细胞核的过程

Escape of DNA from mitochondria to the nucleus in Saccharomyces cerevisiae.

作者信息

Thorsness P E, Fox T D

机构信息

Section of Genetics and Development, Cornell University, Ithaca, New York 14853-2703.

出版信息

Nature. 1990 Jul 26;346(6282):376-9. doi: 10.1038/346376a0.

Abstract

The migration of genetic information from ancestral prokaryotic endosymbionts into eukaryotic nuclei is thought to have had an important role in the evolution of mitochondria and chloroplasts. Here we describe an assay for the detection of movement of DNA between mitochondria and the nucleus in yeast. Because recombinant plasmid DNA replicates after transformation into mitochondria of yeast strains lacking endogenous mitochondrial DNA we were able to propagate the nuclear genetic marker URA3 in mitochondria. As expected, the wild-type URA3 gene in mitochondria failed to complement the uracil auxotrophy (Ura-) caused by a nuclear ura3 mutation. But selection of Ura+ prototrophs from a Ura- strain carrying URA3 on a plasmid in its mitochondria enabled us to detect plasmid movement to the nucleus. Conversely, as the plasmid used also contained the mitochondrial gene COX2 required for respiratory growth, we were able to set up corresponding selections to detect migration of DNA from the nucleus to the mitochondria. Our results show that, in yeast, DNA escapes from mitochondria and appears in the nucleus at a surprisingly high frequency (approximately 2 x 10(-5) per cell per generation). But the rate at which DNA makes the journey in the opposite direction--nucleus to mitochondria--is apparently at least 100,000 times less.

摘要

遗传信息从祖先原核内共生体迁移到真核细胞核被认为在线粒体和叶绿体的进化中起到了重要作用。在此,我们描述了一种检测酵母中线粒体与细胞核之间DNA移动的方法。由于重组质粒DNA在转化到缺乏内源性线粒体DNA的酵母菌株的线粒体后会复制,我们得以在线粒体中繁殖核遗传标记URA3。正如预期的那样,线粒体中的野生型URA3基因无法弥补由核ura3突变导致的尿嘧啶营养缺陷型(Ura-)。但是,从线粒体中携带URA3质粒的Ura-菌株中选择Ura+原养型,使我们能够检测到质粒向细胞核的移动。相反,由于所使用的质粒还包含呼吸生长所需的线粒体基因COX2,我们能够进行相应的选择以检测DNA从细胞核向线粒体的迁移。我们的结果表明,在酵母中,DNA从线粒体逃逸并以惊人的高频率(约每细胞每代2×10^(-5))出现在细胞核中。但是DNA朝相反方向——从细胞核到线粒体——移动的速率显然至少要低100,000倍。

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