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Mutations in the PCNA-binding domain of CDKN1C cause IMAGe syndrome

Nature Genetics volume 44, pages 788–792 (2012)Cite this article

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Abstract

IMAGe syndrome (intrauterine growth restriction, metaphyseal dysplasia, adrenal hypoplasia congenita and genital anomalies) is an undergrowth developmental disorder with life-threatening consequences1. An identity-by-descent analysis in a family with IMAGe syndrome2 identified a 17.2-Mb locus on chromosome 11p15 that segregated in the affected family members. Targeted exon array capture of the disease locus, followed by high-throughput genomic sequencing and validation by dideoxy sequencing, identified missense mutations in the imprinted gene CDKN1C (also known as P57KIP2) in two familial and four unrelated patients. A familial analysis showed an imprinted mode of inheritance in which only maternal transmission of the mutation resulted in IMAGe syndrome. CDKN1C inhibits cell-cycle progression3, and we found that targeted expression of IMAGe-associated CDKN1C mutations in Drosophila caused severe eye growth defects compared to wild-type CDKN1C, suggesting a gain-of-function mechanism. All IMAGe-associated mutations clustered in the PCNA-binding domain of CDKN1C and resulted in loss of PCNA binding, distinguishing them from the mutations of CDKN1C that cause Beckwith-Wiedemann syndrome, an overgrowth syndrome4.

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Figure 1: IBD analysis in a family with IMAGe syndrome.
Figure 2: Localization of IMAGe syndrome–associated mutations in CDKN1C.
Figure 3: Phenotypic validation of IMAGe syndrome–associated mutations in D. melanogaster.
Figure 4: CDKN1C is expressed in the developing human adrenal gland, and IMAGe mutants lose PCNA binding, altering the ubiquitination of CDKN1C.
Figure 5: Missense mutations in the CDK-binding domain and truncating mutations in CDKN1C cause BWS, whereas missense mutations localized to the PCNA-binding domain result in IMAGe syndrome.

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Acknowledgements

This work was funded by the Doris Duke Charitable Foundation and National Institute of Child Health and Human Development RO1HD068138. V.A.A. was supported by the US National Institutes of Health (NIH) 1 F31HD068136 training grant. The human embryonic and fetal material was provided by the Joint Medical Research Council (grant G0700089) and Wellcome Trust (grant GR082557) Human Developmental Biology Resource (www.hdbr.org). J.C.A. was supported by a Wellcome Trust Senior Research Fellowship in Clinical Science (079666). We thank R. Matera (NIH) for kindly providing the pCI-neo-(HA)3-human ubiquitin construct. We thank E.R.B. McCabe for initial crucial support of the adrenal research for E.V. and for providing one of the original IMAGe patient's DNA samples. We thank M. Le Merrer and C. Lecointre, who participated in the initial clinical description of IMAGe.

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Authors and Affiliations

  1. Department of Human Genetics, David Geffen School of Medicine, University of California, Los Angeles, California, USA

    Valerie A Arboleda, Hane Lee, Alice Fleming, Abhik Banerjee, Imilce A Rodriguez-Fernandez, Esteban C Dell'Angelica, Stanley F Nelson, Julian A Martinez-Agosto & Eric Vilain

  2. Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles, California, USA

    Hane Lee & Stanley F Nelson

  3. Developmental Endocrinology Research Group, Clinical and Molecular Genetics Unit, University College London Institute of Child Health, London, UK

    Rahul Parnaik, Bruno Ferraz-de-Souza & John C Achermann

  4. Department of Endocrinology, Laboratory of Medical Investigation (LIM18), University of São Paulo School of Medicine, São Paulo, Brazil

    Bruno Ferraz-de-Souza

  5. Department of Pediatrics, David Geffen School of Medicine, University of California, Los Angeles, California, USA

    Emmanuèle C Délot, Julian A Martinez-Agosto & Eric Vilain

  6. Division of Endocrinology, Hospital de Niños Ricardo Gutierrez, Buenos Aires, Argentina

    Debora Braslavsky & Ignacio Bergadá

  7. Department of Urology, David Geffen School of Medicine, University of California, Los Angeles, California, USA

    Eric Vilain

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  1. Valerie A Arboleda
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Contributions

V.A.A. designed and performed the experiments, analyzed data and wrote the paper. E.V. designed the project, supervised the overall experiments and wrote the paper with V.A.A. H.L. and S.F.N. contributed to design and analysis of the linkage and sequencing data. E.C. Délot contributed to the design of cell-cycle analysis experiments and editing of the manuscript. A.F., E.C. Dell'Angelica and I.A.R.-F. contributed to the nuclear localization experiments and design of the PCNA and ubiquitin assays. D.B. and I.B. clinically assessed and extracted DNA from family A. R.P., B.F.-d.-S. and J.C.A. performed immunofluorescence and RT-PCR experiments. A.B. and J.A.M.-A. performed, analyzed and contributed to the reporting of the Drosophila experiments. All authors discussed the results and implications of the work and commented on the manuscript at various stages.

Corresponding author

Correspondence to Eric Vilain.

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The authors declare no competing financial interests.

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Arboleda, V., Lee, H., Parnaik, R. et al. Mutations in the PCNA-binding domain of CDKN1C cause IMAGe syndrome. Nat Genet 44, 788–792 (2012). https://doi.org/10.1038/ng.2275

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