Figure 4.
In vitro splicing analysis of pre-tRNAs by ARMAN-2 tRNA splicing endonuclease. (A) Pre-tRNAIle (UAU) with 25 nt intron inserted at canonical position (37/38) forming a strict hBHBh′ motif and (B) Pre-tRNACys (GCA) with 13 nt intron located at non-canonical position (59/60) forming a relaxed hBH motif. Pre-tRNAs were incubated with the ARMAN-2 splicing endonuclease for 1 h at different temperatures. Cleaved products (black arrow) were analyzed by 8 M–urea 15% PAGE and stained with SYBR Green II. M, molecular marker; U, uncut product.