The characterization of a novel, putative trypanosome transcription factor

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Description: Trypanosoma brucei, the causative agent of African sleeping sickness, require a 39-nucleotide capped Spliced Leader RNA (SL RNA) trans -spliced onto the 5' end of their messenger RNA (mRNA) for translation. The SL RNA gene is transcribed by RNA Polymerase II and contains the only RNA Polymerase II promoter identified to date in trypanosomes. The interaction between the putative transcription factor Trypanosoma brucei TFIIB-Associated Factor, 49kDa ( Tb TAF49) with the SL RNA gene promoter was studied by promoter pulldown assays. Promoter pulldown assays, using biotinylated DNA mixed with nuclear extracts, were analyzed by Western blotting and the results reveal Tb TAF49 interacts nonspecifically with DNA even when challenged with the presence of high concentrations of NP-40 and Tween-20. Purified GST-TbTAF49 was analyzed by BN-PAGE analysis and gel filtration chromatography to determine the oligomeric state of Tb TAF49. These studies indicate that Tb TAF49 oligomerizes and exists as a multimeric protein.
Language: English
Format: Degree Work