Electronic Thesis and Dissertation Repository

Degree

Master of Science

Program

Biochemistry

Supervisor

Dr. Stanley D. Dunn

2nd Supervisor

Dr. Michael J. Strong

Joint Supervisor

Abstract

Transactive response DNA binding protein, 43 kDa (TDP-43) is 416-residue RNA processing and transport protein, observed in insoluble cytoplasmic aggregates within affected neurons in neurodegenerative diseases. TDP-43 has three domains: the N-terminal (N), RNA-binding (R) and unstructured C-terminal domains (G). Unstructured domains often form intramolecular interactions regulating other domains; our goal was to determine if such an interaction occurs in TDP-43. In Far Western blots, tagged NR was observed to bind to G. A 10 residue C-terminal truncation of G virtually abolished binding and introduction of phosphomimetics at Ser409/Ser410 also reduced binding. Sedimentation velocity ultracentrifugation with tagged NR and G also revealed interaction, observed by a shift in sedimentation coefficients when compared to those of the individual polypeptides. In vivo colocalization studies confirmed a cellular interaction between fluorescently labeled NR and G. This interaction has potential implications for the regulation of TDP-43 and the mechanism of generation of aggregative forms.

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Biochemistry Commons

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