Abstract
Noonan syndrome (NS) is a developmental disorder characterized by short stature, facial dysmorphisms and congenital heart defects. To date, all mutations known to cause NS are dominant, activating mutations in signal transducers of the RAS/mitogen-activated protein kinase (MAPK) pathway. In 25% of cases, however, the genetic cause of NS remains elusive, suggesting that factors other than those involved in the canonical RAS/MAPK pathway may also have a role. Here, we used family-based whole exome sequencing of a case–parent trio and identified a de novo mutation, p.(Arg802His), in A2ML1, which encodes the secreted protease inhibitor α-2-macroglobulin (A2M)-like-1. Subsequent resequencing of A2ML1 in 155 cases with a clinical diagnosis of NS led to the identification of additional mutations in two families, p.(Arg802Leu) and p.(Arg592Leu). Functional characterization of these human A2ML1 mutations in zebrafish showed NS-like developmental defects, including a broad head, blunted face and cardiac malformations. Using the crystal structure of A2M, which is highly homologous to A2ML1, we identified the intramolecular interaction partner of p.Arg802. Mutation of this residue, p.Glu906, induced similar developmental defects in zebrafish, strengthening our conclusion that mutations in A2ML1 cause a disorder clinically related to NS. This is the first report of the involvement of an extracellular factor in a disorder clinically related to RASopathies, providing potential new leads for better understanding of the molecular basis of this family of developmental diseases.
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Acknowledgements
We thank Hanka Venselaar for bioinformatics support in protein modeling and Martina Ruiterkamp-Versteeg, Petra de Vries, Suzanne Keijzers-Vloet and Martine van Zweeden for technical assistance. This work was funded, in part, by a grant from the Research Council for Earth and Life Sciences (ALW 819.02.021) with financial aid from the Netherlands Organization for Scientific Research (NWO) (to JdH), Telethon-Italy (GGP13107 to MT), Fundação para a Ciência e Tecnologia (PTDC/BIM-MEC/0650/2012 to JLC) and PPS5 Consórcio DoIT (ADI – Agência de Inovação to JLC). IPATIMUP is an Associate Laboratory of the Portuguese Ministry of Education and Science and is partially supported by FCT, the Portuguese Foundation for Science and Technology.
Deposition of genetic data
The data obtained in this study are submitted to LOVD, an online gene-centered collection and display of DNA variations (http://databases.lovd.nl/shared/genes).
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Vissers, L., Bonetti, M., Paardekooper Overman, J. et al. Heterozygous germline mutations in A2ML1 are associated with a disorder clinically related to Noonan syndrome. Eur J Hum Genet 23, 317–324 (2015). https://doi.org/10.1038/ejhg.2014.115
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DOI: https://doi.org/10.1038/ejhg.2014.115
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