Figure 2.
tiRNAs can be repaired to full-length tRNAs by RtcB in vitro. Gel-purified tiRNAs were incubated with bacterial RtcB. (A) SYBR Gold staining and (B) Northern blotting for tRNAGly-GCC and tRNASer-GCT. The lengths of both tRNAGly-GCC and tRNASer-GCT are also shown. tRNASer-GCT is longer than tRNAGly-GCC because of its long variable loop. Ligation products are indicated by arrowheads. Red arrowheads indicate the ligation products that showed the same mobility as that of the corresponding mature tRNA in total RNA. Note that two bands were detected by the probe for 5′-side (position 1–21) of tRNASer-GCT, in contrast to a single band detected by the probe targeting 3′-side (position 50–82) of tRNASer-GCT, which suggests that this shorter band (indicated by a blue arrowhead) consists of 5′-tiRNASer-GCT and 3′-tiRNAs derived from non-serine tRNAs.