Project description:Gradient fractions of RNAi of XAC1 (Tb927.7.2780) in Trypanosoma brucei bloodstream forms. RNAi was induced using tetracycline and cell extracts were fractionated into polysomal and monosome-non-ribosome-associated fractions.
Project description: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.
Project description:Surface-exposed proteins of the bloodstream and procyclic forms of T. brucei were biotinylated, affinity purified using streptavidin, and analyzed by LC-MS/MS