Figure 2.
In vitro angiogenin cleavage assays. A, angiogenin digestion pattern of in vitro transcripts of WT (U22) and mutant (MT) (C22) tRNALeu(UUR). Two μg of purified tRNA transcripts were used for the ANG cleavage reaction over time (from 0 to 40 min). Cleavage products of tRNA transcripts were electrophoresed through a denaturing polyacrylamide gel and stained with methylene blue. B, angiogenin digestion pattern of tRNALeu(UUR) purified from control (C47) and mutant (II-5) lymphoblastoid cell lines. Two μg of RNAs were used for the ANG cleavage reaction at various lengths (from 0 to 90 min). Cleavage products of tRNAs were resolved in 15% denaturating polyacrylamide gels with 8 m urea, electroblotted, and hybridized with DIG-labeled oligonucleotide probe specific for the tRNALeu(UUR).