Tandem inverted repeats in mitochondrial DNA of petite mutants of Saccharomyces cerevisiae

Abstract

Denatured mitochondrial DNA (mtDNA) from a grande (wild-type) yeast strain and a series of derived genetically characterized cytoplasmic petite mutants was examined in the electron microscope as DNA-protein monolayers prepared under conditions that permitted little bimolecular renaturation. In the grande and some petite strains, the mtDNA remained predominantly single-stranded. However, in several petite strains, a large proportion of molecules contained double-stranded segments indicative of unimolecular renaturation due to the presence of inverted repeat sequences. The length of the double-stranded segments of strain E41 was compared to the periodicity seen on denaturation maps. A repeat spacing twice the length of the inverted repeats was observed in the denaturation map. Inverted repeat length was similar to contour length of circular mtDNA molecules in this strain. On the basis of these observations most of the mtDNA from petite strain E41 appeared to consist of polymers of tandem inverted repeats interspersed with a small single-stranded "spacer" sequence between the repeat segments. In contrast, petite strain F13 mtDNA had few or no inverted repeats and showed a regular periodicity of 0.14 mum in the denaturation map, similar in length to the 0.13-mum circles present in the isolated mtDNA.