title: A map of 5-methylcytosine residues in Trypanosoma brucei tRNA revealed by sodium bisulfite sequencing.
abstract: In protozoan parasites, there is little information on the presence of covalent RNA modifications which comprise the epitranscriptome. Therefore, we determined if T. brucei tRNA(Asp(GUC)), tRNA(Gly(GCC)), tRNA(Val(AAC)), and tRNA(Tyr(GUA)) contain 5-methylcytosines via RNA bisulfite sequencing. Most tRNAs examined have at least one 5-methylcytosine at the variable region-TψC junction. Only tRNA(Gly(GCC)) displayed methylation of C40 in the anticodon stem, and there was partial methylation at this site. There is no evidence for methylation of C38 in the anticodon loop in the tRNAs analyzed. Analysis of tRNA(Tyr(GUA)) demonstrates that both unspliced and spliced molecules contain C48 methylation, indicating tRNA cytosine methylation can precede tRNA splicing. Overall, our data indicate that T. brucei tRNAs contain 5-methylcytosine residues in some, but potentially not all standard eukaryotic positions. The levels of cytosine methylation of different T. brucei tRNAs vary, suggesting the presence of a mechanism for methylation control.
authors.affiliation: State University of New York at Geneseo, Department of Biology, 1 College Circle, Geneseo, NY 14454, United States. Electronic address: militello@geneseo.edu.
authors.forename: Kevin T
authors.initials: KT
authors.lastname: Militello
authors.affiliation: State University of New York at Geneseo, Department of Biology, 1 College Circle, Geneseo, NY 14454, United States.
authors.forename: Leanne M
authors.initials: LM
authors.lastname: Chen
authors.affiliation: State University of New York at Geneseo, Department of Biology, 1 College Circle, Geneseo, NY 14454, United States.
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Ackerman
authors.affiliation: State University of New York at Geneseo, Department of Biology, 1 College Circle, Geneseo, NY 14454, United States.
authors.forename: Alexandra H
authors.initials: AH
authors.lastname: Mandarano
authors.affiliation: State University of New York at Geneseo, Department of Biology, 1 College Circle, Geneseo, NY 14454, United States.
authors.forename: Erika L
authors.initials: EL
authors.lastname: Valentine
mesh_terms: D044503:5-Methylcytosine; D001483:Base Sequence; D008745:Methylation; D008969:Molecular Sequence Data; D012343:RNA, Transfer / Q000235:genetics*; D017423:Sequence Analysis, RNA / Q000379:methods; D013447:Sulfites; D014346:Trypanosoma brucei brucei / Q000235:genetics*
chemical_list: D013447:Sulfites; D044503:5-Methylcytosine; D012343:RNA, Transfer; C009279:sodium bisulfite
keywords: 5-Methylcytosine; Dnmt2; NSun2; Sodium bisulfite sequencing; tRNA