Variation, transcription and circular RNAs of the mitochondrial gene for subunit I of cytochrome c oxidase.

The gene for subunit I of cytochrome c oxidase, contained within the OX13 region of yeast mitochondrial DNA, is split and shows a remarkable variation in structure, which is strain-dependent. The most complex form so far characterized is that of the Saccharomyces cerevisiae strain KL14-4A, in which nine or possibly ten exons are separated by eight to nine introns. At least four of these are facultative, two being absent from S. cerevisiae strain D273-10B (sequenced by Bonitz et al., 1980) and a further two lacking from the gene in Saccharomyces carlsbergensis. The complexity of the gene in KL14-4A is also reflected in its transcript pattern. RNA blot hybridization with isolated and cloned DNA fragments of the OX13 region permits visualization of more than 60 RNAs, which show overlapping and discontinuous hybridization behaviour. In the less complex strains D273-10B and S. carlsbergensis, this number is 20 and 11, respectively. These RNAs are most likely intermediates in processing events leading to the appearance of the mature messenger RNA for cytochrome c oxidase subunit I, which we identify as a 2100-nucleotide transcript (18SE). Most of the processing events are dependent on mitochondrial protein synthesis and do not constitute a single obligatory processing pathway. Like other yeast mitochondrial mRNAs, the 18 S RNA contains a long, untranslated 5' flanking sequence (approximately 400 nucleotides). One unusual aspect of splicing events involving OX13 transcripts is the accumulation of three of the excised introns as single-stranded RNA circles. These abundant and stable transcripts appear to be covalently closed. The simplest assumption is that they arise as (by)-products of splicing, but secondary ligation events have not been excluded. Their function is as yet unknown.