Usp9X Controls Ankyrin-Repeat Domain Protein Homeostasis during Dendritic Spine Development

Sehyoun Yoon; Euan Parnell; Maria Kasherman; Marc P Forrest; Kristoffer Myczek; Susitha Premarathne; Michelle C Sanchez Vega; Michael Piper; Thomas H J Burne; Lachlan A Jolly; Stephen A Wood; Peter Penzes

doi:10.1016/j.neuron.2019.11.003

Usp9X Controls Ankyrin-Repeat Domain Protein Homeostasis during Dendritic Spine Development

Neuron. 2020 Feb 5;105(3):506-521.e7. doi: 10.1016/j.neuron.2019.11.003. Epub 2019 Dec 5.

Affiliations

¹ Department of Physiology, Northwestern University Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA.
² Griffith Institute for Drug Discovery, Griffith University, Brisbane, QLD 4111, Australia; The School of Biomedical Sciences, The University of Queensland, Brisbane, QLD 4072 Australia.
³ Griffith Institute for Drug Discovery, Griffith University, Brisbane, QLD 4111, Australia.
⁴ Queensland Brain Institute, The University of Queensland, Brisbane, QLD 4072, Australia.
⁵ The School of Biomedical Sciences, The University of Queensland, Brisbane, QLD 4072 Australia; Queensland Brain Institute, The University of Queensland, Brisbane, QLD 4072, Australia.
⁶ Queensland Brain Institute, The University of Queensland, Brisbane, QLD 4072, Australia; Queensland Centre for Mental Health Research, The Park Centre for Mental Health, Wacol, QLD 4076, Australia.
⁷ Robinson Research Institute, School of Medicine, University of Adelaide, Adelaide, SA 5005, Australia.
⁸ Department of Physiology, Northwestern University Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA; Department of Psychiatry and Behavioral Sciences, Northwestern University Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA. Electronic address: p-penzes@northwestern.edu.

Abstract

Variants in the ANK3 gene encoding ankyrin-G are associated with neurodevelopmental disorders, including intellectual disability, autism, schizophrenia, and bipolar disorder. However, no upstream regulators of ankyrin-G at synapses are known. Here, we show that ankyrin-G interacts with Usp9X, a neurodevelopmental-disorder-associated deubiquitinase (DUB). Usp9X phosphorylation enhances their interaction, decreases ankyrin-G polyubiquitination, and stabilizes ankyrin-G to maintain dendritic spine development. In forebrain-specific Usp9X knockout mice (Usp9X^-/Y), ankyrin-G as well as multiple ankyrin-repeat domain (ANKRD)-containing proteins are transiently reduced at 2 but recovered at 12 weeks postnatally. However, reduced cortical spine density in knockouts persists into adulthood. Usp9X^-/Y mice display increase of ankyrin-G ubiquitination and aggregation and hyperactivity. USP9X mutations in patients with intellectual disability and autism ablate its catalytic activity or ankyrin-G interaction. Our data reveal a DUB-dependent mechanism of ANKRD protein homeostasis, the impairment of which only transiently affects ANKRD protein levels but leads to persistent neuronal, behavioral, and clinical abnormalities.

Keywords: ANK; SHANK; ankyrin-G; deubiquitinase; intellectual disability; proximity ligation assay; structured illumination microscopy.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Ankyrin Repeat / physiology*
Cells, Cultured
Dendritic Spines / physiology*
HEK293 Cells
Homeostasis / physiology*
Humans
Male
Mice
Mice, Inbred C57BL
Mice, Knockout
Neurogenesis / physiology
Protein Structure, Secondary
Protein Structure, Tertiary
Proteostasis / physiology*
Ubiquitin Thiolesterase / chemistry
Ubiquitin Thiolesterase / genetics
Ubiquitin Thiolesterase / metabolism*

Substances

Ubiquitin Thiolesterase
Usp9x protein, mouse

Grants and funding

R01 MH107182/MH/NIMH NIH HHS/United States