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TDP-43 Phosphorylation State Regulates Solubility, Localization, and Cellular Toxicity

Tennant, William (2024) TDP-43 Phosphorylation State Regulates Solubility, Localization, and Cellular Toxicity. Doctoral Dissertation, University of Pittsburgh. (Unpublished)

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Abstract

The hyperphosphorylated and insoluble TDP-43 inclusions are a pathological hallmark of several neurodegenerative disorders such as amyotrophic lateral sclerosis (ALS), frontal temporal dementia (FTD), and Alzheimer's Disease (AD). The mechanism by which phosphorylation regulates TDP-43 physiological functions and its contribution to the observed neuropathology remain unclear. To assess this, we generated recombinant TDP-43 proteins that mimic the hyperphosphorylated and hypophosphorylated condition to study the effect of TDP-43 phosphorylation of the LCD on phase separation, cellular localization, and cytotoxicity. Our studies show that both hyper- and hypo- phosphorylation mimetics promote TDP-43 mislocalization to the cytoplasm , enhance the propensity and stability of the TDP-43 C-terminal domain protein droplets, and disrupt phase separation. However, in all models it is only the phosphodeficient condition that reduces TDP-43 solubility and promotes the formation of cytotoxic aggregates. Furthermore, phosphodeficient TDP-43 induces the sequestration of endogenous TDP-43 to these insoluble aggregates that ultimately diminish TDP-43 splicing capacity. Taken together, our efforts provide further evidence that insoluble, cytoplasmic TDP-43 inclusions are cytotoxic and that phosphorylation might actually play a protective role, inhibiting the formation of insoluble, toxic, cytoplasmic TDP-43 aggregates.

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Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Tennant, Williamwmt7@pitt.eduwmt70000-0002-8555-7881
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Thesis AdvisorDonnelly, Christopherchrisdonnelly@pitt.educjdon250000-0002-2383-9015
Committee ChairChu, CTctc4@pitt.eduCTC40000-0002-5052-8271
Committee MemberBerman, Sarahbermans@upmc.edu0000-0002-5096-4962
Committee MemberTalbott, Evelyneot1@pitt.eduEOT1
Committee MemberHwang, Hun-wayhunway.hwang@pitt.edu0000-0003-0522-1577
Committee MemberKolarcik, Christiclk39@pitt.educlk39
Date: 16 September 2024
Date Type: Publication
Defense Date: 9 February 2024
Approval Date: 16 September 2024
Submission Date: 4 April 2024
Access Restriction: 1 year -- Restrict access to University of Pittsburgh for a period of 1 year.
Number of Pages: 108
Institution: University of Pittsburgh
Schools and Programs: School of Medicine > Cellular and Molecular Pathology
Degree: PhD - Doctor of Philosophy
Thesis Type: Doctoral Dissertation
Refereed: Yes
Uncontrolled Keywords: ALS, TDP-43, Phosphorylation, Liquid-liquid phase separation, neurodegenerative disease, RNA binding proteins, post-translational modifications
Date Deposited: 16 Sep 2024 19:09
Last Modified: 16 Sep 2024 19:09
URI: http://d-scholarship.pitt.edu/id/eprint/46014

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