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Trypanosoma brucei is a protozoan parasite that causes human African Trypanosomiasis (sleeping sickness) in people and nagana in cattle, both of which are fatal without treatment. This parasite is injected into the host through the bite of the tsetse fly and is able to evade the host’s immune response due to changes in its major surface antigen, variant surface glycoproteins (VSGs). This constant switching prevents the host from making a single antibody that can recognize the antigen and eliminate the parasite. VSG expression sites have been found to be near the telomeres of Trypanosoma brucei, and studies from our lab have shown that a telomere protein called TbRAP1 is essential for regulating VSG silencing. In order to better understand the mechanisms of how TbRAP1 silences VSGs, we aim to identify proteins that interact with TbRAP1. We have done a yeast 2-hybrid screen using the TbRAP1 fulllength protein as bait previously, but TbRAP1 itself has a weak transcription activation function that leads to identification of false-positive candidates. TbRAP1 protein has several functional domains, among which the BRCT domain has a weak transcription activation activity while others do not. In the current study, we aim to perform a yeast 2-hybrid screen using TbRAP1-aa426-761 fragment as bait. Without the basal transcription activation activity from the TbRAP1 BRCT domain, we will be able to identify true interacting factors of the TbRAP1-aa426-761 fragment. One initial screen has been performed, resulting in 10 candidates. We are currently performing another screen, hoping to identify more interacting candidates.
College of Sciences and Health Professions
Biological, Geological, and Environmental Sciences
Radzin, Annelise and Beran, Elizabeth, "Screen for interacting factors for Trypanosoma brucei telomere protein RAP1" (2018). Undergraduate Research Posters 2018. 19.