FIGURE 8.

Processing and degradation of M. genitalium tRNA₁ ^Gly precursor. Pre-tRNA₁ ^Gly with a mature 5′-end and a 21-nt 3′-trailer was uniformly labeled with ³²P and was treated with RNases as described in Materials and Methods. RNA products were detected by autoradiography after separation on an 8% denaturing polyacrylamide gel. (Lane 1) A [³²P]-labeled RNA ladder (RNA Decade Markers; Ambion) was used as the size marker. (Lanes 2,3) In addition, 78-nt and 74-nt RNA transcripts containing the same sequence as tRNA₁ ^Gly were used as size markers. These markers were generated using PCR templates from the tRNA gene, starting from the 5′-end of the tRNA and ending at the 3′-end (for 74 nt) or 4 nt downstream (for 78 nt). Pre-tRNA₁ ^Gly was treated by buffer (containing 10 μM ZnCl₂), EcR, MgR, or EcII for 30 min at 37°C in the presence of various concentrations of MgCl₂ as indicated on the top of each lane. (P) The size of the precursor is 95 nt. (M) The size of mature tRNA₁ ^Gly is 74 nt.