Transcriptional initiation and 5' termini of yeast mitochondrial RNA

We have used vaccinia virus guanylyltransferase to label polyphosphate-terminated yeast mitochondrial RNAs in vitro with [alpha-32P]GTP. Hybridization of RNA labeled in vitro indicates the presence of multiple transcriptional initiation sites in both grande and petite mitochondrial genomes. Agarose/urea gel electrophoresis of capped RNA suggests the existence of a precursor to the small (14 S) rRNA. In contrast, direct examination of the large (21 S) rRNA by partial ribonuclease T1 digestion reveals a complete lack of processing of the 5' end of the primary transcript of this RNA.