Date of Award
Chemistry and Biochemistry
Erich G. Chapman, Ph.D.
Aggregation, Amyotrophic lateral sclerosis, Neurodegenerative disorders, Phosphomimetic, S48E, TDP-43, TAR DNA-binding protein 43
Trans-activation response (TAR) DNA-binding protein 43 (TDP-43) is a natively dimeric 414-residue protein that is encoded by the human TARDBP 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 Escherichia coli 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 in vitro. We show that the addition of RNA/DNA to the S48E mutant does not have profound effects on aggregation attenuation under in vitro aggregation conditions.
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Toro, Nicole, "Characterization of a Phosphomimetic Mutant of the ALS Associated Protein TDP-43" (2019). Electronic Theses and Dissertations. 1628.
Received from ProQuest
Biochemistry, Molecular biology