Date of Award

1-1-2019

Document Type

Masters Thesis

Degree Name

M.S.

Organizational Unit

College of Natual Science and Mathematics, Chemistry and Biochemistry

First Advisor

Erich G. Chapman, Ph.D.

Keywords

Aggregation, Amyotrophic lateral sclerosis, Neurodegenerative disorders, Phosphomimetic, S48E, TDP-43, TAR DNA-binding protein 43

Abstract

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.

Publication Statement

Copyright is held by the author. User is responsible for all copyright compliance.

Rights Holder

Nicole Toro

Provenance

Received from ProQuest

File Format

application/pdf

Language

en

File Size

72 p.

Discipline

Biochemistry, Molecular biology

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