The large repertoire of 2’-O-methylation guided by C/D snoRNAs on Trypanosoma brucei rRNA

K. Shanmugha Rajan, Yinzhou Zhu, Katerina Adler, Tirza Doniger, Smadar Cohen-Chalamish, Ankita Srivastava, Moran Shalev-Benami, Donna Matzov, Ron Unger, Christian Tschudi, Arthur Günzl, Gordon G. Carmichael, Shulamit Michaeli

Research output: Contribution to journalArticlepeer-review

Abstract

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 2ʹ-O-methylations (Nms) on trypanosome rRNA using three high-throughput sequencing methods; RibOxi-seq, RiboMeth-seq and 2ʹ-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.

Original languageEnglish
Pages (from-to)1018-1039
Number of pages22
JournalRNA Biology
Volume17
Issue number7
DOIs
StatePublished - 2 Jul 2020

Keywords

  • 2ʹ-OMe-seq
  • RibOxi-seq
  • RiboMeth-seq
  • Trypanosomes
  • rRNA
  • snoRNA

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

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