Identification and functional implications of pseudouridine RNA modification on small noncoding RNAs in the mammalian pathogen Trypanosoma brucei

Trypanosoma brucei, the parasite that causes sleeping sickness, cycles between an insect and a mammalian host. However, the effect of RNA modifications such as pseudouridinylation on its ability to survive in these two different host environments is unclear. Here, two genome-wide approaches were applied for mapping pseudouridinylation sites (psi s) on small nucleolar RNA (snoRNA), 7SL RNA, vault RNA, and tRNAs from T. brucei. We show using HydraPsiSeq and RiboMeth-seq that the psi on C/D snoRNA guiding 2'-O-methylation increased the efficiency of the guided modification on its target, rRNA. We found differential levels of psi s on these noncoding RNAs in the two life stages (insect host and mammalian host) of the parasite. Furthermore, tRNA isoform abundance and psi modifications were characterized in these two life stages demonstrating stage-specific regulation. We conclude that the differential psi modifications identified here may contribute to modulating the function of noncoding RNAs involved in rRNA processing, rRNA modification, protein synthesis, and protein trans-location during cycling of the parasite between its two hosts.

Automated summary

The effect of RNA modifications on the survival ability of Trypanosoma brucei, the parasite causing sleeping sickness, in insect and mammalian hosts is unclear. This study mapped pseudouridinylation sites on different RNAs in the parasite using two genome-wide approaches. The researchers found differential levels of pseudouridinylation on noncoding RNAs in the two life stages of the parasite, suggesting a potential role in regulating important biological processes.