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A hypofunctional PAX1 mutation causes autosomal recessively inherited otofaciocervical syndrome

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An Erratum to this article was published on 25 July 2013

Abstract

Otofaciocervical syndrome (OFCS) is an autosomal recessively inherited disorder characterized by facial dysmorphism, external ear anomalies with preauricular pits and hearing impairment, branchial cysts or fistulas, anomalies of the vertebrae and the shoulder girdle, and mild intellectual disability. In a large consanguineous family with OFCS from Turkey, we performed whole-exome sequencing (WES) of a single pooled DNA sample of four affected individuals. Filtering for variants with a percentage of alternate reads ≥90 % and a coverage of at least five reads identified only a single novel homozygous variant, c.497G>T, located in PAX1 that co-segregated with the disease in the family. PAX1 encodes a transcription factor with a critical role in pattern formation during embryogenesis in vertebrates. The mutation is predicted to substitute the glycine at position 166 to valine (p.G166V) within the highly conserved paired-box domain of the PAX1 protein. We performed a dual luciferase reporter assay to examine the transactivation of a regulatory sequence in the Nkx3-2 promoter region, which is a direct target of mouse Pax1 transcriptional regulation. We observed a significantly reduced transactivation in HEK293T cells overexpressing Pax1G157V in comparison to Pax1WT expressing cells, indicating a reduced DNA-binding affinity of the mutant protein. Taken together, our results show that the strategy of pooling DNA is a powerful, cost-effective application for WES in consanguineous families and establish PAX1 as a new disease-causing gene for OFCS and as part of the EYA-DACH-SIX-PAX network, important in early embryogenesis.

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Acknowledgments

We are grateful to all family members who participated in this study, to Esther Milz for excellent technical assistance, to Andrea Pannes for her technical advice in slow-down PCR, to Simon von Ameln for cochlea preparation, and to Karin Boss for critically reading the manuscript. This work was supported by the German Federal Ministry of Education and Research (BMBF) by grant number 01GM1211A (E-RARE network CRANIRARE-2) to B.W.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Institute of Human Genetics, University Medical Faculty, University of Cologne, Kerpener Str. 34, 50931, Cologne, Germany

    Esther Pohl, Ayca Aykut, Filippo Beleggia, Katharina Keupp, Gökhan Yigit & Bernd Wollnik

  2. Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany

    Esther Pohl, Filippo Beleggia, Katharina Keupp, Gökhan Yigit & Bernd Wollnik

  3. Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany

    Esther Pohl, Filippo Beleggia, Katharina Keupp, Gökhan Yigit & Bernd Wollnik

  4. Department of Medical Genetics, Faculty of Medicine, Ege University, Izmir, Turkey

    Ayca Aykut, Emin Karaca, Burak Durmaz, Esra Arslan & Ferda Özkinay

  5. Department of Ophthalmology, Faculty of Medicine, Ege University, Izmir, Turkey

    Melis Palamar Onay

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  1. Esther Pohl
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Correspondence to Bernd Wollnik.

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Pohl, E., Aykut, A., Beleggia, F. et al. A hypofunctional PAX1 mutation causes autosomal recessively inherited otofaciocervical syndrome. Hum Genet 132, 1311–1320 (2013). https://doi.org/10.1007/s00439-013-1337-9

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  • DOI: https://doi.org/10.1007/s00439-013-1337-9

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