Skip to main content
. 2003 Apr 1;31(7):1904–1912. doi: 10.1093/nar/gkg282

Figure 5.

Figure 5

Solution secondary structure probing of wild-type and G4309A tRNAIle. tRNAs [wild-type and G4309A in (A) and (B), respectively] were labeled at their 5′ ends and cleaved with NaHCO3 to produce the alkali ladder (lane 1, designated AL at bottom) and probed with ribonuclease T1 at 1 and 2.5 × 10–3 U/μl under semi-denaturing conditions (lanes 2 and 3, respectively; SD T1). tRNAs re-folded under native conditions (see Materials and Methods) were probed with T1 at 0, 1 and 2.5 × 10–3 U/µl (lanes 4–6; N T1), with ribonuclease S1 at 0.55 and 1.37 U/µl (lanes 7–8) and with ribonuclease V1 at 1 and 2.5 × 10–6 U/µl (lanes 9–10). Numbers on the left designate T1 cleavages at Gs observed in the semi-denaturing T1 lanes. Designations on the right indicate regions and nucleotide numbers corresponding to V1 cleavages, and numbers enclosed in ovals correspond to regions that can be placed on the canonical tRNA structure (see Fig. 6). The arrow on the left and the bar at the right of (B) indicate the position of the G4309A substitution and a corresponding region of altered S1 and V1 sensitivity in this mutant (see Fig. 6).