Figure 4.

Determination of the exact cleavage site by recombinant 3′ tRNases, ELAC2, YKR079C and ELAC1. The unlabeled 5′ cleavage products generated in the in vitro 3′-tRNase assays were 3′-end-labeled with [5′-³²P]pCp and T4 RNA ligase, gel-purified and subjected to chemical RNA sequencing (24). The sequencing reactions were analyzed on a 20% polyacrylamide–8 M urea gel. The 3′-terminal nucleotide G corresponds to the discriminator.