Lack of site-specific recombination between mitochondrial genomes of petite mutants of yeast

Abstract

Previous work from our laboratory showed that mitochondrial (mt) genomes, with tandem repeat units, from spontaneous, cytoplasmic petite mutants of Saccharomyces cerevisiae do not exhibit site-specific recombination in petite x petite crosses [Rayko et al., Gene 63 (1988) 213-226]. Here, we have extended and confirmed these observations by studying other crosses of petites with tandem repeat units, as well as crosses in which one of the parents was, instead, an unstable petite, a-15/4/1, having a palindromic mt genome. In no case was site-specific recombination of the parental mt genomes observed. Progeny cells harbored mt genomes derived from either one or both of the two parents, as shown by analysis of restriction fragments. In the case of biparental inheritance, extensive subcloning of the diploids showed that this was due to a persistent heteroplasmic state and not to intermolecular recombination. The 'new' restriction fragments present in the mt DNA from some diploids were shown to be derived from the unstable parental genome, a-15/4/1, by a secondary excision process. Lack of site-specific recombination is, therefore, not only a feature of crosses involving petite genomes made up of tandem repeat units, but also of crosses in which one parental genome consists of inverted repeats and frequently originates secondary petite genomes formed by tandem repeats. Previous reports of mt recombination in petite mutants are discussed in light of these results.