Journal
RNA BIOLOGY
Volume 17, Issue 7, Pages 1018-1039Publisher
TAYLOR & FRANCIS INC
DOI: 10.1080/15476286.2020.1750842
Keywords
Trypanosomes; RibOxi-seq; RiboMeth-seq; 2MODIFIER LETTER PRIME-OMe-seq; rRNA; snoRNA
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Funding
- Israel-US Binational Science Foundation (BSF)
- I-core programme of the Planning and budgeting committee
- Israel Science Foundation [1796/12]
- NIH [R01 AI028798, AI110325, R21 AI142149, R21 MH110955]
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The parasite Trypanosoma brucei cycles between insect and mammalian hosts, and is the causative agent of sleeping sickness. Here, we performed genome-wide mapping of 2MODIFIER LETTER PRIME-O-methylations (Nms) on trypanosome rRNA using three high-throughput sequencing methods; RibOxi-seq, RiboMeth-seq and 2MODIFIER LETTER PRIME-OMe-seq. This is the first study using three genome-wide mapping approaches on rRNA from the same species showing the discrepancy among the methods. RibOxi-seq detects all the sites, but RiboMeth-seq is the only method to evaluate the level of a single Nm site. The sequencing revealed at least ninety-nine Nms guided by eighty-five snoRNAs among these thirty-eight Nms are trypanosome specific sites. We present the sequence and target of the C/D snoRNAs guiding on rRNA. This is the highest number of Nms detected to date on rRNA of a single cell parasite. Based on RiboMeth-seq, several Nm sites were found to be differentially regulated at the two stages of the parasite life cycle, the insect procyclic form (PCF) versus the bloodstream form (BSF) in the mammalian host.
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