A maturase-like subunit of the sequence-specific endonuclease endo.SceI from yeast mitochondria

Abstract

Some yeast strains possess a sequence-specific endonuclease, Endo.SceI, which is a heterodimeric enzyme localized in mitochondria. The larger subunit (75 kDa) of Endo.SceI, encoded by a nuclear gene (ENS1), is transported from the cytosol into the mitochondria. In this study, we determined the partial amino acid sequence of the smaller subunit (50 kDa) of Endo.SceI. The determined sequence matched well the partial sequence deduced from a mitochondrial open reading frame (RF3). The RF3 locus is known to exhibit polymorphism since this reading frame in some yeast strains is supposed to encode a maturase-like protein, whereas in other strains, the frame is interrupted by GC clusters, which thus break the frame. Southern blot analysis of various yeast strains showed that the continuity of RF3 is correlated with the presence of Endo.SceI activity. These data indicate that the continuous RF3 sequence is a functional gene (ENS2) coding for the smaller subunit of Endo.SceI. The results of cytoduction, by which the continuous RF3 sequence was transferred into a yeast strain lacking mitochondrial DNA, confirmed this conclusion. This study suggests the involvement of Endo.SceI in genetic recombination of mitochondrial DNA.