Mutations at a single codon in Mad homology 2 domain of SMAD4 cause Myhre syndrome

Abstract

Myhre syndrome (MIM 139210) is a developmental disorder characterized by short stature, short hands and feet, facial dysmorphism, muscular hypertrophy, deafness and cognitive delay. Using exome sequencing of individuals with Myhre syndrome, we identified SMAD4 as a candidate gene that contributes to this syndrome on the basis of its pivotal role in the bone morphogenetic pathway (BMP) and transforming growth factor (TGF)-β signaling. We identified three distinct heterozygous missense SMAD4 mutations affecting the codon for Ile500 in 11 individuals with Myhre syndrome. All three mutations are located in the region of SMAD4 encoding the Mad homology 2 (MH2) domain near the site of monoubiquitination at Lys519, and we found a defect in SMAD4 ubiquitination in fibroblasts from affected individuals. We also observed decreased expression of downstream TGF-β target genes, supporting the idea of impaired TGF-β–mediated transcriptional control in individuals with Myhre syndrome.

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Figure 1: Clinical and radiological manifestations of individuals with Myhre syndrome.
Figure 2: Functional consequences of SMAD4 mutations in fibroblasts from individuals with Myhre syndrome.
Figure 3: Levels of phosphorylated SMAD proteins in skin fibroblasts from individuals with Myhre syndrome and age- and passage-matched controls.
Figure 4: Cellular localization of phosphorylated SMAD proteins.
Figure 5: Expression analysis of TGF-β– and BMP-driven target genes in fibroblasts from control and case subjects.

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Acknowledgements

We are grateful to the individuals with Myhre syndrome and their families for their participation in this study. We also thank the following physicians for the management of the affected individuals: D. Doummar, R. McGowan, P. Picco and M. Whiteford. This research was supported by the French National Research Agency (ANR; R09183KS to V.C.-D.).

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Contributions

C.L.G. designed the experiments, analyzed the exome sequencing data, performed protein blot analysis and wrote the manuscript. C.M. performed Sanger sequencing analysis. A. Abhyankar and J.-L.C. performed exome capture. W.L.G. performed quantitative RT-PCR analysis. V.S. performed three-dimensional structure analysis. A. Afenjar, A.D., M.d.R., D.H., S.J., S.M., M.S., J.T. and A.V. provided clinical data. A.M. wrote the manuscript. V.C.-D. provided clinical data, analyzed the exome sequencing data, oversaw all aspects of the research and wrote the manuscript.

Corresponding author

Correspondence to Valérie Cormier-Daire.

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The authors declare no competing financial interests.

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Supplementary Figures 1 and 2 and Supplementary Tables 1–4. (PDF 307 kb)

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Le Goff, C., Mahaut, C., Abhyankar, A. et al. Mutations at a single codon in Mad homology 2 domain of SMAD4 cause Myhre syndrome. Nat Genet 44, 85–88 (2012). https://doi.org/10.1038/ng.1016

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