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Stabilizing mutations of KLHL24 ubiquitin ligase cause loss of keratin 14 and human skin fragility

Nature Genetics volume 48, pages 1508–1516 (2016)Cite this article

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Abstract

Skin integrity is essential for protection from external stress and trauma. Defects in structural proteins such as keratins cause skin fragility, epitomized by epidermolysis bullosa (EB), a life-threatening disorder. Here we show that dominant mutations of KLHL24, encoding a cullin 3–RBX1 ubiquitin ligase substrate receptor, cause EB. We have identified start-codon mutations in the KLHL24 gene in five patients with EB. These mutations lead to truncated KLHL24 protein lacking the initial 28 amino acids (KLHL24-ΔN28). KLHL24-ΔN28 is more stable than its wild-type counterpart owing to abolished autoubiquitination. We have further identified keratin 14 (KRT14) as a KLHL24 substrate and found that KLHL24-ΔN28 induces excessive ubiquitination and degradation of KRT14. Using a knock-in mouse model, we have confirmed that the Klhl24 mutations lead to stabilized Klhl24-ΔN28 and cause Krt14 degradation. Our findings identify a new disease-causing mechanism due to dysregulation of autoubiquitination and open new avenues for the treatment of related disorders.

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Figure 1: Pedigree, clinical features and skin pathology in an inherited skin fragility disorder.
Figure 2: KLHL24 start-codon mutations identified in the five patients result in an N-terminally truncated protein.
Figure 3: Removal of the first 28 residues of KLHL24 increases protein stability as a result of abolished autoubiquitination.
Figure 4: KRT14 is a ubiquitination substrate of KLHL24 and is excessively ubiquitinated and degraded by KLHL24-ΔN28.
Figure 5: Klhl24 mRNA levels are upregulated during mouse keratinocyte differentiation, and mutation of its start codon causes stabilization of KLHL24 and loss of Krt14 in a knock-in mouse model.
Figure 6: Schematic of the disease-causing mechanism of KLHL24 mutations.

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Article 24 September 2020

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Acknowledgements

We thank J. Jin, G. Ou, L. Yu, G. Xu and N. Zheng for advice and support, A. Elia, M. Emanuele and S. Elledge for critical reading of the manuscript, and all the patients and their families for participating in this study. This work was supported by the National Natural Science Foundation of China (grants 81201220 to Z.L., 81271744 to Y.Y. and 31670777 to X.T.), the Young Thousand Talents Program of China to X.T., China National Funds for Distinguished Young Scientists (grant 81425020 to Y.Y.), China National Funds for Excellent Young Scientists (grant 81522037 to Z.L.) and the Beijing Nova Program (grant Z151100000315044 to Z.L.).

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  1. Zhimiao Lin, Shuo Li and Cheng Feng: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Dermatology, Peking University First Hospital, Beijing Key Laboratory of Molecular Diagnosis on Dermatoses, Beijing, China

    Zhimiao Lin, Cheng Feng, Huijun Wang, Jing Zhang, Dingfang Bu, Xintong Wang & Yong Yang

  2. School of Pharmaceutical Sciences, Center for Infectious Disease Research, School of Medicine, Tsinghua University, Tsinghua-Peking Center for Life Sciences, Beijing, China

    Shuo Li, Shang Yang, Danhui Ma, Mengting Gou, Tengjiang Zhang, Xiaohui Kong & Xu Tan

  3. Peking-Tsinghua Center for Life Sciences, Beijing, China

    Huijun Wang, Jing Zhang & Yong Yang

  4. Department of Dermatology, Department of Human Molecular Genetics and Biochemistry, Tel Aviv Sourasky Medical Centre, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel

    Ofer Sarig, Gilly Padalon-Brauch, Dan Vodo & Eli Sprecher

  5. Laboratory of Electron Microscopy, Peking University First Hospital, Beijing, China

    Yali Ren

  6. BGI-Shenzhen, Shenzhen, China

    Lanlan Dai, Hankui Liu & Jianguo Zhang

  7. National Institute of Biological Sciences, Beijing, China

    Fei Li & Ting Chen

  8. Laboratory Animal Facility, Peking University First Hospital, Beijing, China

    Yongyan Hu

  9. Institutes of Biomedical Sciences, Shanghai Medical College of Fudan University, Shanghai, China

    Feng Zhou

  10. MOE Key Laboratory of Bioinformatics, School of Life Sciences and Centre of Biomedical Analysis, Tsinghua University, Beijing, China

    Haiteng Deng

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  1. Zhimiao Lin
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Contributions

Z.L., S.L., C.F., S.Y., H.W., D.M., Jing Zhang, M.G., X.K., D.B., T.Z., Y.R., X.W. and Y.H. performed the experiments under the supervision of X.T. and Y.Y. O.S., E.S., F.L., T.C., G.P.-B. and D.V. contributed critical reagents. L.D., H.L. and Jianguo Zhang performed the exome sequencing and data analysis. H.D. and F.Z. contributed to the proteomics data analysis. X.T., C.F., S.L., Z.L. and Y.Y. wrote the manuscript.

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Correspondence to Yong Yang or Xu Tan.

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Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–17 and Supplementary Tables 1–3. (PDF 1846 kb)

Supplementary Table 4

Sequences of primers, sgRNAs, siRNAs and probes used in the study. (XLSX 39 kb)

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Lin, Z., Li, S., Feng, C. et al. Stabilizing mutations of KLHL24 ubiquitin ligase cause loss of keratin 14 and human skin fragility. Nat Genet 48, 1508–1516 (2016). https://doi.org/10.1038/ng.3701

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