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.
","abstract_html":"<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>","abstract_has_math":false,"creators":["Toro, Nicole"],"institution":null,"degree_name":"M.S.","degree_level":"Masters Thesis","degree_discipline":null,"degree_department":null,"school":null,"contributors":["Erich G. Chapman, Ph.D."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2019,"date_issued":"2019-01-01T08:00:00Z","date_published":"2019-01-01T08:00:00Z","updated_at":"2026-07-24T02:02:11Z","subjects":["Aggregation","Amyotrophic lateral sclerosis","Neurodegenerative disorders","Phosphomimetic","S48E","TDP-43","TAR DNA-binding protein 43","Life Sciences","Neuroscience and Neurobiology"],"languages":["en"],"rights":["Copyright is held by the author. User is responsible for all copyright compliance.
"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://digitalcommons.du.edu/etd/1628","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Erich G. Chapman, Ph.D."]},{"key":"dc:creator","label":"Author","values":["Toro, Nicole"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2019-08-02T07:00:00Z"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Masters Thesis"]},{"key":"thesis:degree_name","label":"Degree Name","values":["M.S."]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Aggregation","Amyotrophic lateral sclerosis","Neurodegenerative disorders","Phosphomimetic","S48E","TDP-43","TAR DNA-binding protein 43","Life Sciences","Neuroscience and Neurobiology"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright is held by the author. User is responsible for all copyright compliance.
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"]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Characterization of a Phosphomimetic Mutant of the ALS Associated Protein TDP-43"]}]}],"canonical_facts":{"dc:contributor":["Erich G. Chapman, Ph.D."],"dc:creator":["Toro, Nicole"],"dc:date.available":["2019-08-02T07:00:00Z"],"dc:description.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.
"],"dc:format":["application/pdf"],"dc:identifier":["https://digitalcommons.du.edu/etd/1628"],"dc:language":["en"],"dc:rights":["Copyright is held by the author. User is responsible for all copyright compliance.
"],"dc:subject":["Aggregation","Amyotrophic lateral sclerosis","Neurodegenerative disorders","Phosphomimetic","S48E","TDP-43","TAR DNA-binding protein 43","Life Sciences","Neuroscience and Neurobiology"],"dc:title":["Characterization of a Phosphomimetic Mutant of the ALS Associated Protein TDP-43"],"thesis:degree_level":["Masters Thesis"],"thesis:degree_name":["M.S."]},"updated_at":"2026-07-24T02:02:11Z"}