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Mutations in PLK4, encoding a master regulator of centriole biogenesis, cause microcephaly, growth failure and retinopathy

Nature Genetics volume 46, pages 1283–1292 (2014)Cite this article

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Abstract

Centrioles are essential for ciliogenesis. However, mutations in centriole biogenesis genes have been reported in primary microcephaly and Seckel syndrome, disorders without the hallmark clinical features of ciliopathies. Here we identify mutations in the genes encoding PLK4 kinase, a master regulator of centriole duplication, and its substrate TUBGCP6 in individuals with microcephalic primordial dwarfism and additional congenital anomalies, including retinopathy, thereby extending the human phenotypic spectrum associated with centriole dysfunction. Furthermore, we establish that different levels of impaired PLK4 activity result in growth and cilia phenotypes, providing a mechanism by which microcephaly disorders can occur with or without ciliopathic features.

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Figure 1: Individuals with mutations in PLK4 or TUBGCP6 exhibit extreme microcephaly and short stature.
Figure 2: Consequences of PLK4 mutations on the transcript and protein.
Figure 3: PLK4 mutations impair PLK4 activity in centriole biogenesis, resulting in reduced centriole number in patient-derived cells.
Figure 4: Depletion of plk4 causes dwarfism in zebrafish.
Figure 5: Impaired mitosis leads to growth retardation in plk4-morphant zebrafish.
Figure 6: plk4-morphant zebrafish display retinal defects due to reduced cilia number.
Figure 7: The growth failure and ciliopathy phenotypes are separable in a dose-dependent manner.

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Acknowledgements

We thank the families and clinicians for their involvement and participation; P. Mills, T. Hurd, M. Bettencourt-Dias and M. Reijns for commenting on the manuscript; N. Hastie, D. Fitzpatrick and J. Livingston for helpful discussions; C. Janke (Institut Curie) for his kind gift of the GT335 antibody; E. Freyer for assistance with FACS analysis; P. Gautier for bioinformatics; P. Carroll and A. Vickers for technical assistance; the IGMM core sequencing service; the IGMM imaging facility for assistance with microscopy; E. Patton and the IGMM fish facility for advice and zebrafish technical assistance; E. Liston and the DNA Resource Centre at SickKids for sample processing; A. Pearce and E. Maher (Cytogenetics Laboratory, South East Scotland Genetics Service) for technical advice; G. Hahn (University Hospital Carl Gustav Carus) for her second opinion on the MRI data; and N. Dalibor and E. Kirst (CCG) for their expert technical assistance. This work was supported by funding from the MRC, the Lister Institute for Preventative Medicine and the European Research Council (ERC, 281847) (A.P.J.), Medical Research Scotland (L.S.B.), the National Institute for Health Research Moorfields Eye Hospital Biomedical Research Centre (A.T.M.), Köln Fortune (M.S.H.) and CMMC (P.N. and A.A.N.).

Author information

Author notes

  1. Carol-Anne Martin, Ilyas Ahmad, Anna Klingseisen and Muhammad Sajid Hussain: These authors contributed equally to this work.

Authors and Affiliations

  1. Medical Research Council (MRC) Human Genetics Unit, Institute of Genetics and Molecular Medicine (IGMM), University of Edinburgh, Edinburgh, UK

    Carol-Anne Martin, Anna Klingseisen, Louise S Bicknell, Andrea Leitch, Jennie E Murray, David Hunt, James Ding, Margaret E Harley, Patricia Heyn & Andrew P Jackson

  2. Cologne Center for Genomics (CCG), University of Cologne, Cologne, Germany

    Ilyas Ahmad, Muhammad Sajid Hussain, Gudrun Nürnberg, Mohammad Reza Toliat, Holger Thiele, Janine Altmüller, Wolfgang Höhne & Peter Nürnberg

  3. Institute of Biochemistry I, Medical Faculty, University of Cologne, Cologne, Germany

    Ilyas Ahmad, Muhammad Sajid Hussain, Rolf Müller & Angelika Anna Noegel

  4. Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Faisalabad, Pakistan

    Ilyas Ahmad, Muhammad Sajid Hussain, Fawad Khan, Zafar Ali & Shahid Mahmood Baig

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

    Muhammad Sajid Hussain, Angelika Anna Noegel & Peter Nürnberg

  6. Division of Human Genetics, Innsbruck Medical University, Innsbruck, Austria

    Sigrid Tinschert

  7. Institute for Clinical Genetics, Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany

    Sigrid Tinschert

  8. Cytogenetics Laboratory, South East of Scotland Genetics Service, Western General Hospital, Edinburgh, UK

    Charlotte Keith

  9. Cell Cycle Control and Carcinogenesis, German Cancer Research Center (DKFZ), Heidelberg, Germany

    Ingrid Hoffmann

  10. Department of Genetics, INSERM U781, Université Paris Descartes, Sorbonne Paris Cité, Hopital Necker, Assistance Publique–Hôpitaux de Paris (AP-HP), Paris, France

    Valérie Cormier Daire

  11. CARGO and IGMA Hôpitaux Universitaires de Strasbourg, INSERM U1112, Université de Strasbourg, Strasbourg, France

    Hélène Dollfus

  12. Division of Clinical and Metabolic Genetics, Department of Paediatrics, The Hospital for Sick Children and University of Toronto, Toronto, Ontario, Canada

    Lucie Dupuis & Roberto Mendoza-Londono

  13. Unit of Human Developmental Genetics, Institut Pasteur, Paris, France

    Anu Bashamboo & Kenneth McElreavey

  14. Najmabadi Pathology and Genetics Center, Tehran, Iran

    Ariana Kariminejad

  15. University College London (UCL) Institute of Ophthalmology, London, UK

    Anthony T Moore

  16. Moorfields Eye Hospital, London, UK

    Anthony T Moore

  17. Southwest Thames Regional Genetics Service, St. George's Hospital Medical School, London, UK

    Anand Saggar

  18. Casey Eye Institute, Oregon Health and Science University, Portland, Oregon, USA

    Catie Schlechter & Richard Weleber

  19. Institute of Human Genetics, University of Cologne, Cologne, Germany

    Janine Altmüller

  20. Wellcome Trust Sanger Institute, Cambridge, UK,

    Matthew E Hurles

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

    Angelika Anna Noegel & Peter Nürnberg

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  1. Carol-Anne Martin
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  2. Ilyas Ahmad
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Contributions

P.N., H.T., J.A., M.S.H., A.B., K.M., M.E. Hurles, J.E.M. and L.S.B. performed exome sequencing and analysis. L.S.B., C.-A.M., J.E.M., M.R.T., I.A., M.S.H. and G.N. performed sequencing, genotyping, linkage analysis and other molecular genetics experiments. C.-A.M., A.L., C.K., M.E. Harley, I.A., M.S.H., R.M., A.A.N. and I.H. designed and performed the cell biology experiments. A. Klingseisen designed and performed the zebrafish experiments with help from A.L., C.-A.M., J.D. and P.H. W.H. performed structural analysis. D.H., F.K., Z.A., S.T., V.C.-D., H.D., L.D., A. Kariminejad, R.M.-L., A.T.M., A.S., C.S., R.W. and S.M.B. ascertained subjects, obtained samples and/or assisted with phenotypic analysis and clinical studies. C.-A.M. and A.P.J. wrote the manuscript with help from P.N., A. Klingseisen and L.S.B. The study was planned and supervised by P.N. and A.P.J.

Corresponding authors

Correspondence to Peter Nürnberg or Andrew P Jackson.

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Competing interests

P.N. is a founder, CEO and shareholder of ATLAS Biolabs. ATLAS Biolabs is a service provider for genomic analyses.

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Martin, CA., Ahmad, I., Klingseisen, A. et al. Mutations in PLK4, encoding a master regulator of centriole biogenesis, cause microcephaly, growth failure and retinopathy. Nat Genet 46, 1283–1292 (2014). https://doi.org/10.1038/ng.3122

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