Characterization of a Phosphomimetic Mutant of the ALS Associated Protein TDP-43

<p>Trans-activation response (TAR) DNA-binding protein 43 (TDP-43) is a natively dimeric 414-residue protein that is encoded by the human <em>TARDBP</em> gene that has important implications in the pathogenesis of the neurodegenerative disorders ALS, FTD, and CTE. TDP-43 has been found hyperphosphorylated and ubiquitinated in the aggregates of the affected neurons of these diseases. The discovery of the presence of TDP-43 positive inclusions in brain matter of patients with CTE has made repetitive brain injury a possible environmental stimulus for aggregation in TDP-43 proteinopathies. We expand upon the hypothesis that TDP-43 readily aggregates under agitation conditions and that the addition of poly-TG repeats to TDP-43 in aggregation conditions attenuates its aggregation propensity. We expressed a recombinant S48E phosphomimetic mutation of TDP-43 with an N-terminal GFP fluorescent tag in <em>Escherichia coli</em> and induced aggregation by agitation. We examined the extent to which different DNA/RNAs and differing stoichiometric concentrations of these nucleic acids affected the aggregation<em> in vitro</em>. We show that the addition of RNA/DNA to the S48E mutant does not have profound effects on aggregation attenuation under <em>in vitro</em> aggregation conditions.</p>

Name thesis:degree_name

M.S.

Level thesis:degree_level

Masters Thesis

Year dc:date.available

2019

Author dc:creator

• Toro, Nicole

Contributors dc:contributor

• Erich G. Chapman, Ph.D.

• Aggregation
• Amyotrophic lateral sclerosis
• Neurodegenerative disorders
• Phosphomimetic
• S48E
• TDP-43
• TAR DNA-binding protein 43
• Life Sciences
• Neuroscience and Neurobiology

Statement dc:rights

• <p>Copyright is held by the author. User is responsible for all copyright compliance.</p>

Language dc:language

en

Repository record dc:identifier

https://digitalcommons.du.edu/etd/1628

OAI identifier oai:identifier

oai:digitalcommons.du.edu:etd-2628