Escape of DNA from mitochondria to the nucleus in Saccharomyces cerevisiae

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^1–6. Here we describe an assay for the detection of movement of DNA between mitochondria and the nucleus in yeast. Because recom-binant plasmid DNA replicates after transformation into mitochondria of yeast strains lacking endogenous mitochondrial DNA⁷ 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⁺ pro-totrophs 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 ( ˜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.