T.brucei ncRNA Interactome
(A) Schematic presentation of the ncRNA interactome identification protocol. The scheme illustrates the in vivo AMT-psoralen UV cross-linking treatment, extract preparation, fractionation, RNA extraction, mild fragmentation, ligation, and the RNA library preparation methodology.
(B) Enrichment of intermolecular RNA-RNA hybrids in ncRNA interactome upon ligation. The results show that the percentage of chimeric RNA increase from negligible to nearly 4% following ligation (left) and indicate the percentage of intermolecular cross-links in two replicates (right).
(C) Ciros plot representing the known interactions of U snRNA in T. brucei following ligation of cross-linked RNAs.
(D) Chimeric snoRNA-pre rRNA. The potential for base-pairing of TB10Cs1C4 with the pre-rRNA, and the coverage of ligation products between these RNAs in RPKM across the pre-rRNA are presented. An example of a chimeric RNA sequence and its target snoRNA is shown, demonstrating that ligation between the RNAs takes place close to the interaction domain. The different domains of the pre-rRNA are indicated.