ADAM17 variant causes hair loss via ubiquitin ligase TRIM47–mediated degradation
Xiaoxiao Wang, Chaolan Pan, Luyao Zheng, Jianbo Wang, Quan Zou, Peiyi Sun, Kaili Zhou, Anqi Zhao, Qiaoyu Cao, Wei He, Yumeng Wang, Ruhong Cheng, Zhirong Yao, Si Zhang, Hui Zhang, Ming Li
Xiaoxiao Wang, Chaolan Pan, Luyao Zheng, Jianbo Wang, Quan Zou, Peiyi Sun, Kaili Zhou, Anqi Zhao, Qiaoyu Cao, Wei He, Yumeng Wang, Ruhong Cheng, Zhirong Yao, Si Zhang, Hui Zhang, Ming Li
View: Text | PDF
Research Article Dermatology Genetics

ADAM17 variant causes hair loss via ubiquitin ligase TRIM47–mediated degradation

Abstract

Hypotrichosis is a genetic disorder characterized by a diffuse and progressive loss of scalp and/or body hair. Nonetheless, the causative genes for several affected individuals remain elusive, and the underlying mechanisms have yet to be fully elucidated. Here, we discovered a dominant variant in a disintegrin and a metalloproteinase domain 17 (ADAM17) gene caused hypotrichosis with woolly hair. Adam17 (p.D647N) knockin mice mimicked the hair abnormality in patients. ADAM17 (p.D647N) mutation led to hair follicle stem cell (HFSC) exhaustion and caused abnormal hair follicles, ultimately resulting in alopecia. Mechanistic studies revealed that ADAM17 binds directly to E3 ubiquitin ligase tripartite motif-containing protein 47 (TRIM47). ADAM17 variant enhanced the association between ADAM17 and TRIM47, leading to an increase in ubiquitination and subsequent degradation of ADAM17 protein. Furthermore, reduced ADAM17 protein expression affected the Notch signaling pathway, impairing the activation, proliferation, and differentiation of HFSCs during hair follicle regeneration. Overexpression of Notch intracellular domain rescued the reduced proliferation ability caused by Adam17 variant in primary fibroblast cells.

Authors

Xiaoxiao Wang, Chaolan Pan, Luyao Zheng, Jianbo Wang, Quan Zou, Peiyi Sun, Kaili Zhou, Anqi Zhao, Qiaoyu Cao, Wei He, Yumeng Wang, Ruhong Cheng, Zhirong Yao, Si Zhang, Hui Zhang, Ming Li

×

Figure 5

TRIM47 is identified as a specific E3 ubiquitin ligase of ADAM17.

Options: View larger image (or click on image) Download as PowerPoint
TRIM47 is identified as a specific E3 ubiquitin ligase of ADAM17. (A) Li...
(A) Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of wild-type and mutant ADAM17 binding proteins. (B) Peptide spectrum match score ratios for the proteins identified by mass spectrometry. (C) ADAM17 interacts with TRIM47 in HaCaT cells, and their association was enhanced by the ADAM17 (p.D647N) mutation. (D) The direct interaction between ADAM17 and TRIM47 was validated by pulldown assay. (E) Endogenous Adam17 binds to Trim47 in the epidermal tissue lysates obtained from mice, and Adam17 (p.D647N) mutation enhances their association. (F) Confocal immunofluorescence revealed a colocalization (yellow) of ADAM17 (red) and TRIM47 (green) in HaCaT cells, and ADAM17 (p.D647N) mutation enhanced their colocalization. Scale bars, 8 μm (first 3 images); 1 μm (fourth image). (G) Confocal immunofluorescence revealed a colocalization (yellow) of Adam17 (red) and Trim47 (green) in primary cultured mouse skin fibroblasts. Scale bars, 10 μm (first 3 images); 1 μm (fourth image). (H) Adam17 and Trim47 colocalized in both IRS and ORS of hair follicles. The white arrows indicated the colocalization (yellow) of Trim47 (green) and Adam17 (red). Scale bars, 40 μm (first 4 images); 20 μm (fifth image). (I) Knockdown of TRIM47 impeded the proteasomal degradation of ADAM17. Left panel: Representative immunoblot images of the ADAM17 and TRIM47 protein levels during CHX chase assays. Right panel: quantification of immunoblotting results corresponding to the left panel. (n = 3 biological replicates.) (J) The 3-dimensional structure of ADAM17/TRIM47 complex in stereo. Right upper panel: essential amino acids of ADAM17 (blue) that polar contacted TRIM47 (green) were depicted. Right lower panel: residues of TRIM47 (green) which polar contacted to ADAM17 (blue) were depicted. All experiments were repeated 3 times. Results were expressed as mean ± SD; *P < 0.05; unpaired 2-tailed t test (I).