A novel ALS-associated variant in UBQLN4 regulates motor axon morphogenesis
- Northwestern University Feinberg School of Medicine, Ann & Robert H Lurie Children's Hospital of Chicago, United States
- The Les Turner ALS Research and Patient Center, Northwestern University Feinberg School of Medicine, United States
Figures
Figure 1
The UBQLN4 c.269A>C (p.D90A) variant identified in a familial ALS case.
(A) Pedigree of a family with ALS. The proband (III3, arrow) had disease onset at 55 years of age, with disease duration of 22 months. Her mother (II3) died of ALS at 62 years of age without clear …
Figure 2 with 2 supplements
Expression of UBQLN4D90A results in motor axon branching abnormalities in vitro and in vivo.
(A) Representative images of primary mouse spinal motor neurons transfected with pCAG-GFP alone, or co-transfected with UBQLN4-WT or UBQLN4D90A. Scale bar: 20 μm. (B) Representative images of …
Figure 2—figure supplement 1
Cultured primary motor neurons express the motor neuron marker Islet1.
(A) Representative images of primary mouse spinal cord neurons transfected with pCAG-GFP and UBQLN4-WT or UBQLN4D90A, stained with Islet1 (red) and DAPI (blue). Scale bar: 20 μm.
Figure 2—figure supplement 2
UBQLN4-WT and UBQLN4D90A are expressed at similar levels in primary mouse motor neurons and in zebrafish embryos.
(A) Representative images of primary mouse spinal cord neurons transfected with pCAG-GFP and Flag-tagged UBQLN4-WT or UBQLN4D90A, stained with anti-Flag antibody (red) and DAPI (blue). Scale bar: 20 …
Figure 3
Expression of UBQLN4D90A impairs proteasomal degradation and results in beta-catenin accumulation.
(A) Representative images of NSC-34 cells transfected with UbG67V-GFP alone, or co-transfected with UBQLN4-WT or UBQLN4D90A. DAPI staining is shown in blue. Scale bar: 100 μm. (B) Quantification of …
Figure 4
UBQLN4D90A-induced phenotypes are rescued by beta-catenin inhibition.
(A) Representative images of primary mouse neurons transfected with pCAG-GFP and UBQLN4-WT or UBQLN4D90A, treated with 0.1 μM quercetin or DMSO. Cells are stained for beta-catenin. Scale bar: 20 μm. …
Figure 5
Schematic model illustrating proposed roles for wild-type (A) and ALS-associated UBQLN4D90A (B) in motor axon morphogenesis.
(A) Wild-type UBQLN4 associates with beta-catenin through its UBA domain, and with the proteasome through its UBL domain. These interactions allow for the degradation of beta-catenin, which in turn …
Additional files
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Supplementary file 1
Human UBQLN4 sequencing primers.
- https://doi.org/10.7554/eLife.25453.010
