Processing of transcripts of a dimeric tRNA gene in yeast uses the nuclease responsible for maturation of the 3′ termini upon 5 S and 37 S precursor rRNAs

The rna82 mutation of Saccharomyces cerevisiae inactivates an RNA processing activity responsible for maturation of 3'-terminal sequences upon 5 S and 37 S ribosomal RNA precursors. This study describes a difference in the processing of transcripts of an S. cerevisiae dimeric tRNA gene (tRNA(arg)-tRNA(Asp) in RNA polymerase III in vitro transcription extracts prepared from rna82 and wild-type cells. The mutant extract accumulated additional processing intermediates containing tRNA(Arg) sequences as compared to the extract from wild-type cells. The structure of these intermediates revealed a defect in removal of the 10 nucleotides left 3' to the tRNA(Arg) sequence by the RNase P cleavage immediately 5' to tRNA(Asp). This is the first demonstration of a mutational defect affecting maturation of 3' sequences upon a eukaryotic tRNA precursor.