TblncRNA-23, a long non-coding RNA transcribed by RNA polymerase I, regulates developmental changes in Trypanosoma brucei

Abstract

Abstract The protozoan parasite Trypanosoma brucei undergoes a complex life cycle, moving between its mammalian host and the blood-feeding tsetse fly vector. The two major surface proteins expressed by procyclic forms in the insect midgut, EP and GPEET procyclin, are transcribed from a polycistronic transcription unit by RNA polymerase I. Here we identify a long non-coding RNA, TblncRNA-23, that is encoded between the two procyclin genes. TblncRNA-23 localizes to the nucleolus and also associates with polysomes. Overexpression of TblncRNA-23 and its down regulation by RNAi or knockout (KO) identify EP and GPEET mRNAs as targets, among other mRNAs that changed abundance in the transition from early to late procyclic forms and from procylic to the metacylic forms, suggesting its role in regulating gene expression which accomapines or dictates of the parasite transitions within in its insect host. TblncRNA-23 interacts with its substrates via base-pairing using different domains. Purification of TblncRNA-23-associated proteins by RaPID identifies hundreds of proteins, including proteins translated from its target mRNAs, suggesting its association with translating ribosomes. Early and late procyclic forms differ in their social motility (SoMo) capabilities, which is essential for migration away from the insect midgut to enable parasite transmission. Overexpression of TblncRNA-23 results in hypermotility, whereas KO compromises this capacity, suggesting a regulatory role in SoMo. Moreover, silencing of the RNA abrogates the ability of the parasite to transform from procylic to the metacyclic forms affecting the parasite’s potential to cycle between its hosts.