Fig. 4. Substrate scope of C12orf29.

RNA oligonucleotides that were used for intramolecular ligation resulting in cyclisation are schematically depicted. The ³²P-labelled 5′-ends were depicted in red. The radioactive oligonucleotides were resolved by denaturing PAGE and analysed by phosphorimaging. a, Investigation of the impact of the nucleobase composition at the 5′- and 3′- termini at the ligation site on the ligation efficiency of C12orf29 (representative images of n = 3). The depicted constructs were incubated under the same conditions for various time points. Most efficient ligation was observed when purines were at the ligation site. b Investigation of the impact of the length of the 5′-overhang on the ligation efficiency of C12orf29 (representative images of n = 3). c Impact of single site mutations on RNA ligase efficiency of C12orf29. Graph bars represent mean ligation efficiencies ± SEM and hollow circles represent individual data points for n = 3 biological replicates. All oligonucleotide sequences are provided in Supplementary Table 1. Source data are provided as a Source Data file.