Electronic Thesis and Dissertation Repository

Degree

Master of Science

Program

Biochemistry

Supervisor(s)

Dr. Stanley D. Dunn and Dr. Michael J. Strong

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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