Torsins are essential members of the AAA+ (ATPases associated with a variety of cellular activities) superfamily and have been implicated in protein quality control, modulation of membrane morphology, and nuclear envelope dynamics. Four different Torsin proteins are known in humans. For years Torsins were thought to lack ATPase activity. Correspondingly, the precise cellular functions and the mechanistic roles of Torsin ATPases remained largely elusive. This gap in our knowledge hinders a comprehensive understanding of the etiology of congenital disorders caused by mutations in the Torsin system, including the severe movement disorder DYT1 dystonia. Importantly, our recent work has revealed that Torsins are ATPases whose activity requires LAP1 or LULL1, which are membrane-spanning cofactors that associate with Torsins to form a composite, membrane-spanning machine. Advances in our functional and mechanistic understanding of Torsin ATPases will require (i) definition of the structure and dynamics of the membrane-bound Torsin/cofactor assembly, (ii) identification of the cellular targets of the Torsin/cofactor machinery, and (iii) functional dissection of Torsin/cofactor action on substrates. In this proposal, we will capitalize on our established proteoliposome system to investigate the structural dynamics of full-length Torsin assembly with and without its cofactors via cryo-electron microscopy (Aim 1). We will utilize a novel methodology to overcome the poor solubility of Torsin substrates, allowing for substrate identification via a subtractive proteomic approach (Aim 2). The substrates, and their dependence on Torsin/cofactor action, will be analyzed in a cellular context, using genetic deletions in concert with different imaging techniques, as well as in reconstituted systems (Aim 3). The elucidation of Torsin function and mechanism -as well as its dysfunction resulting from disease-associated mutations- will enable the development of targeted therapies for the treatment of Torsin-related movement disorders, which are the most common inherited movement disorders known.

Public Health Relevance

DYT1 dystonia is a severe congenital movement disorder caused by a mutation in TorsinA, an essential membrane-associated molecular motor. At present, the development of effective therapies is severely hampered by our incomplete understanding of TorsinA function. We will combine structural, biochemical, and cell biological approaches to delineate the cellular functions of TorsinA and investigate on a cellular and structural level how these functions are perturbed by dystonia-causing mutations. Results stemming from our proposed studies will close a critical gap in our fundamental understanding of the Torsin machinery and lay the groundwork to develop novel therapeutic strategies for the treatment of DYT1 dystonia.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM114401-04
Application #
9694252
Study Section
Membrane Biology and Protein Processing Study Section (MBPP)
Program Officer
Flicker, Paula F
Project Start
2016-06-01
Project End
2020-05-30
Budget Start
2019-06-01
Budget End
2020-05-30
Support Year
4
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Yale University
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Chase, Anna R; Laudermilch, Ethan; Wang, Jimin et al. (2017) Dynamic functional assembly of the Torsin AAA+ ATPase and its modulation by LAP1. Mol Biol Cell 28:2765-2772
Chase, Anna R; Laudermilch, Ethan; Schlieker, Christian (2017) Torsin ATPases: Harnessing Dynamic Instability for Function. Front Mol Biosci 4:29
Laudermilch, Ethan; Tsai, Pei-Ling; Graham, Morven et al. (2016) Dissecting Torsin/cofactor function at the nuclear envelope: a genetic study. Mol Biol Cell 27:3964-3971
Turner, Elizabeth M; Schlieker, Christian (2016) Pelger-Hu√ęt anomaly and Greenberg skeletal dysplasia: LBR-associated diseases of cholesterol metabolism. Rare Dis 4:e1241363