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Insight into the epigenetic regulation of gene expression in the bloodstream and procyclic forms of Trypanosoma brucei through the involvement of G-Quadruplexes

ABSTRACT: The study investigates the epigenetic role of G-quadruplexes (G4s) in the differentiation of Trypanosoma brucei brucei, specifically between its bloodstream form (BF) and procyclic form (PC). An in silico analysis identified 115,126 potential G4 sequences (PQSs) across the genome, with 63% having a high probability of G4 formation. These PQSs are unevenly distributed, with notable enrichment in regions associated with antigenic variation, such as variant surface glycoproteins and bloodstream expression sites. The differential distribution of PQSs correlates with regions of high densities of differentially expressed genes, suggesting a regulatory role in morphological transitions. The study assessed the effects of G4-ligands (AQ1, Pt-TTPY, and pyridostatin) on gene expression, revealing significant downregulation of DEGs and highlighting their therapeutic potential. Functional analysis using Gene Ontology indicated that G4-ligands impact various biological processes differently in BF and PC forms. These findings underscore the epigenetic complexity of T. brucei and the potential of G4 structures as key regulators of gene expression and differentiation, offering novel insights into therapeutic strategies targeting the parasite's adaptive mechanisms.

ORGANISM(S): Trypanosoma brucei

PROVIDER: GSE275674 | GEO | 2024/08/31

REPOSITORIES: GEO

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Project description:Trypanosoma brucei subspecies infect humans and animals in sub-Saharan Africa. This early diverging eukaryote shows many novel features in basic biological processes, including the use of polycistronic transcription to generate all protein-coding mRNAs. Therefore we hypothesized that translational control provides a means to tune gene expression during parasite development in mammalian and fly hosts. We used ribosome profiling to examine genome-wide protein production in animal-derived slender bloodstream forms and cultured procyclic (insect midgut) forms. About one-third of all CDSs showed statistically significant regulation of protein production between the two stages. Of these, more than two-thirds showed a change in translation efficiency, but few were controlled by this parameter alone. Ribosomal proteins were translated poorly, especially in animal-derived parasites. A disproportionate number of metabolic enzymes were up-regulated at the mRNA level in procyclic forms, as were variant surface glycoproteins in bloodstream forms. Comparison with cultured bloodstream forms from another strain identified stage-specific changes in protein production that transcend strain and growth conditions. Genes with upstream ORFs had a lower mean translation efficiency but no evidence was seen for involvement of these ORFs in stage-regulation. Ribosome profiling revealed that differences in the production of specific proteins in T. brucei slender bloodstream and procyclic forms are more common than anticipated from analysis of mRNA abundance. While in vivo and in vitro derived slender bloodstream forms of different strains are more similar to one another than to procyclic forms, they showed numerous differences at both the mRNA and protein production level. Ribosome profiling and mRNA libraries were constructed in triplicate from in vitro PCF and in vivo BF lifestages of theT. brucei Treu927 and in vitro T. brucei Lister427, to evaluate role of translational gene regulation

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Insight into the epigenetic regulation of gene expression in the bloodstream and procyclic forms of Trypanosoma brucei through the involvement of G-Quadruplexes

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