Structure and transcription of the mitochondrial genome in heteroplasmic strains ofSaccharomyces cerevisiae

Saccharomyces cerevisiae strain FF1210-6C/170 is respiratory deficient due to a mutation of the penultimate base of the mitochondrial tRNA(Asp) gene. We have identified a number of progeny from this strain which have reverted to respiratory sufficiency by the excision and tandem amplification of a small region of the mitochondrial (mt) DNA carrying the tRNA(Asp) gene, while also maintaining the full-length mtDNA. We have studied the structure of the mtDNA and mitochondrial transcription in a number of these heteroplasmic strains. The exact site of the recombination involved in the excision of the repeating unit of the amplified mtDNA has been determined for five of the revertants. Recombination occurs between identical sequences 4-13 base pairs in length. Each of the different repeating units of the amplified DNA retains an active promoter which has been moved to a site just upstream of the tRNA(Asp) gene by the excision/amplification. Transcripts from the heteroplasmic strains have been characterized to determine the sites of mitochondrial RNA termini. We find that in addition to the 5' and 3' processing of the tRNAs, many of the transcripts terminate at a position about 300 base pairs downstream of the gene for tRNA(Asp). We also find that 3' processing of tRNA(Asp) precursors is absent in petite strains which lack 5' processing indicating that 5' processing of tRNA(Asp) may be a prerequisite for 3' processing in this mutant.