Abstract
Background: The contrasting physiological environments of Trypanosoma brucei procyclic (insect vector) and bloodstream (mammalian host) forms necessitates deployment of different molecular processes and, therefore, changes in protein expression. Transcriptional regulation is unusual in T. bruceibecause the arrangement of genes is polycistronic; however, genes which are transcribed together are subsequently cleaved into separate mRNAs by trans-splicing. Following pre-mRNA processing, the regulation of mature mRNA stability is a tightly controlled cellular process. While many stage-specific transcripts have been identified, previous studies using RNA-seq suggest that changes in overall transcript level do not necessarily reflect the abundance of the corresponding protein. Methods: To better understand the regulation of gene expression in T. brucei, we performed a bioinformatic analysis of RNA-seq on total, sub-polysomal, and polysomal mRNA samples. We further cross-referenced our dataset with a previously published proteomics dataset to identify new protein coding sequences. Results: Our analyses showed that several long non-coding RNAs are more abundant in the sub-polysome samples, which possibly implicates them in regulating cellular differentiation in T. brucei. We also improved the annotation of the T.brucei genome by identifying new putative protein coding transcripts that were confirmed by mass spectrometry data. Conclusions: Several long non-coding RNAs are more abundant in the sub-polysome cellular fractions and might pay a role in the regulation of gene expression. We hope that these data will be of wide general interest, as well as being of specific value to researchers studying gene regulation expression and life stage transitions in T. brucei.
Highlights
Trypanosoma brucei, a protozoan parasite transmitted by the tsetse fly, causes human African trypanosomiasis (HAT) and nagana in cattle[1]
The high protein content of polysomes allows them to be separated throughout a sucrose gradient according to the number of ribosomes attached to the mRNA (Figure 1)
While RBP10 itself was not found in our sub-polysome enriched transcript list, the long non-coding RNA (lncRNA) (KS17gene_1749a) which is predicted to be at the 5’ end of RBP10 may have a similar regulatory function as the grumpy lncRNA transcript
Summary
Trypanosoma brucei, a protozoan parasite transmitted by the tsetse fly, causes human African trypanosomiasis (HAT) and nagana in cattle[1]. Some of the slender BSF parasites differentiate into non-replicative stumpy forms in the bloodstream and these are pre-adapted for transformation into replicating procyclic form in the testse vector midgut. Results: Our analyses showed that several long non-coding RNAs are more abundant in the sub-polysome samples, which possibly implicates them in regulating cellular differentiation in T. brucei. Conclusions: Several long non-coding RNAs are more abundant in the sub-polysome cellular fractions and might pay a role in the regulation of gene expression. We hope that these data will be of wide general interest, as well as being of specific value to researchers studying gene regulation expression and life stage transitions in T. brucei
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