N6-methyladenosine (m6A) is the most prevalent mRNA modification in higher eukaryotes. Recent studies suggest that m6A has a regulatory role in mRNA degradation and translation initiation or efficiency, involving in cell fate determination in yeast, plants, and stem cells of mammalian. Trypanosoma brucei (T. brucei) regulates gene expression through post-transcriptional fashion, which heavily relies on mRNA cis-motifs. However, internal mRNA modification in T. brucei has not been reported yet. Here we found m6A modification is abundant in T. brucei and presented a transcriptome wide methylome of m6A in both life stages of T. brucei. We identified 355 and 95 peaks in procyclic form and blood stream form trypanosomes respectively. A consensus motif of CAU was shared in both life stages of T. brucei. mRNA abundance of m6A-containing genes is higher in procyclic form and tend to be down-regulated in bloodstream form trypanosomes. Furthermore, m6A-containing transcripts harbor relative longer half-lives, and are enriched in pathways of cell morphology and movement in procyclic form trypanosomes. By m6A-containing RNA pulldown in both life stages, we identified TRRM2 as a potential m6A reader in T. brucei. Uncovering the m6A methylome and its binding proteins may provide a new post-transcriptional regulatory pathway in T. brucei. (C) 2018 Elsevier Inc. All rights reserved.