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A new reporter cell line for studies with proteasome inhibitors in Trypanosoma brucei.

Accepted version
Peer-reviewed

Type

Article

Change log

Authors

Moura, Danielle MN 
de Melo Neto, Osvaldo P 

Abstract

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.

Description

Keywords

Lactacystin, Proteasome, Trypanosoma brucei, Ubiquitin, Cell Line, Drug Evaluation, Preclinical, Genes, Reporter, Green Fluorescent Proteins, Humans, Proteasome Endopeptidase Complex, Proteasome Inhibitors, Trypanocidal Agents, Trypanosoma brucei brucei, Trypanosomiasis, African, Ubiquitin

Journal Title

Mol Biochem Parasitol

Conference Name

Journal ISSN

0166-6851
1872-9428

Volume Title

227

Publisher

Elsevier BV
Sponsorship
Wellcome Trust (085956/Z/08/Z)