Using Tetracysteine-Tagged TDP-43 with a Biarsenical Dye To Monitor Real-Time Trafficking in a Cell Model of Amyotrophic Lateral Sclerosis.
Ng, Janice SW
Hanspal, Maya A
Barros, Teresa P
Wilson, Mark R
Dobson, Christopher M
American Chemical Society (ACS)
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Ng, J. S., Hanspal, M. A., Matharu, N., Barros, T. P., Esbjörner, E. K., Wilson, M. R., Yerbury, J. J., et al. (2019). Using Tetracysteine-Tagged TDP-43 with a Biarsenical Dye To Monitor Real-Time Trafficking in a Cell Model of Amyotrophic Lateral Sclerosis.. Biochemistry, 58 (39), 4086-4095. https://doi.org/10.1021/acs.biochem.9b00592
TAR DNA-binding protein 43 (TDP-43) has been identified as the major constituent of the proteinaceous inclusions that are characteristic of most forms of amyotrophic lateral sclerosis (ALS) and ubiquitin positive frontotemporal lobar degeneration (FTLD). Wild type (WT) TDP-43 inclusions are a pathological hallmark of >95% of patients with sporadic ALS and of the majority of familial ALS cases, and they are also found in a significant proportion of FTLD cases. ALS is the most common form of motor neurone disease, characterised by progressive weakness and muscular wasting, and typically leads to death within a few years of diagnosis. To determine how the translocation and misfolding of TDP-43 contributes to ALS pathogenicity, it is crucial to define the dynamic behaviour of this protein within the cellular environment. It is therefore necessary to develop cell models that allow the location of the protein to be defined. We report the use of TDP-43 with a tetracysteine tag for visualisation using fluorogenic biarsenical compounds, and show that this model displays features of ALS observed in other cell models. We also demonstrate that this labelling procedure enables live-cell imaging of the translocation of the protein from the nucleus into the cytosol.
Cell Line, Tumor, Cell Nucleus, Cytosol, Humans, Amyotrophic Lateral Sclerosis, Organometallic Compounds, Cysteine, Fluoresceins, DNA-Binding Proteins, Fluorescent Dyes, Electroporation, Transfection, Amino Acid Sequence, Protein Transport, Kinetics, Sequence Tagged Sites, Models, Biological, Frontotemporal Lobar Degeneration, Time-Lapse Imaging, Optical Imaging
Wellcome Trust (094425/Z/10/Z)
External DOI: https://doi.org/10.1021/acs.biochem.9b00592
This record's URL: https://www.repository.cam.ac.uk/handle/1810/296687
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