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- W3173660716 abstract "Trypanosoma brucei is the causative agent of African Sleeping Sickness. The regulation of gene expression in T. brucei primarily occurs at the posttranscriptional level, indicating that RNA processing and modification events are extremely important for the parasite's life cycle. Yet, there is little information regarding the collection of covalent RNA modifications that comprise the epitranscriptome in this organism or any protozoan parasite. We focused on cytosine base modifications of tRNAs, as tRNAs are well known to contain a range of modified RNA bases, including 5-methylcytosine. Small RNAs were isolated from insect-stage T. brucei parasites and unmodified cytosines were converted to uracils using sodium bisulfite treatment. Sodium bisulfite-treated small RNAs were analyzed by RNAseq. To confirm the RNAseq data, sodium bisulfite-treated RNA samples were amplified by RT-PCR from either total RNA or small RNA fractions, and analyzed by digestion with HpaII or via Sanger sequencing. Overall, our data indicate that tRNA molecules contain between zero to four 5-methylcytosine residues. The most common location of 5-methylcytosine in T. brucei tRNA is at the junction between the variable region and TψC arm at positions C48, C49, and C50, although different tRNAs have different numbers of 5-methylcytosines in this region. There was little evidence for methylation of other sites in the tRNA, including C38 methylation in the anticodon loop. Overall, our data indicated that T. brucei tRNAs contain 5-methylcytosine at some, but not all standard eukaryotic positions, and the levels of 5-methylcytosine vary in different tRNA molecules." @default.
- W3173660716 created "2021-07-05" @default.
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- W3173660716 date "2015-04-01" @default.
- W3173660716 modified "2023-10-15" @default.
- W3173660716 title "The tRNA Methylome of Trypanosoma brucei , the Causative Agent of African Sleeping Sickness" @default.
- W3173660716 doi "https://doi.org/10.1096/fasebj.29.1_supplement.711.8" @default.
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