A new reporter cell line for studies with proteasome inhibitors in Trypanosoma brucei.
Moura, Danielle MN
de Melo Neto, Osvaldo P
Mol Biochem Parasitol
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Moura, D. M., de Melo Neto, O. P., & Carrington, M. (2019). A new reporter cell line for studies with proteasome inhibitors in Trypanosoma brucei.. Mol Biochem Parasitol, 227 15-18. https://doi.org/10.1016/j.molbiopara.2018.11.001
A Trypanosoma brucei cell line is described that produces a visual readout of proteasome activity. The cell line contains an integrated transgene encoding an ubiquitin-green fluorescent protein (GFP) fusion polypeptide responsive to the addition of proteasome inhibitors. A modified version of T. brucei ubiquitin unable to be recognized by deubiquitinases (UbG76V) was fused to eGFP and constitutively expressed. The fusion protein is unstable but addition of the proteasome inhibitor lactacystin stabilizes it and leads to visually detectable GFP. This cell line can be widely used to monitor the efficiency of inhibitor treatment through detection of GFP accumulation in studies involving proteasome-mediated proteolysis, screening of proteasome inhibitors or other events related to the ubiquitin-proteasome pathway.
Cell Line, Humans, Trypanosoma brucei brucei, Trypanosomiasis, African, Proteasome Endopeptidase Complex, Green Fluorescent Proteins, Ubiquitin, Trypanocidal Agents, Drug Evaluation, Preclinical, Genes, Reporter, Proteasome Inhibitors
Wellcome Trust (085956/Z/08/Z)
External DOI: https://doi.org/10.1016/j.molbiopara.2018.11.001
This record's URL: https://www.repository.cam.ac.uk/handle/1810/287078