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Analysis of cilia dysfunction phenotypes in zebrafish embryos depleted of Origin recognition complex factors

European Journal of Human Genetics (2019) | Download Citation

Abstract

Meier–Gorlin syndrome (MGS) is a rare, congenital primordial microcephalic dwarfism disorder. MGS is caused by genetic variants of components of the origin recognition complex (ORC) consisting of ORC1–6 and the pre-replication complex, which together enable origin firing and hence genome replication. In addition, ORC1 has previously been shown to play a role in ciliogenesis. Here, we extend this work and investigate the function of ORC1 and two other members of the complex on cilia at an organismal level. Knockdown experiments in zebrafish confirmed the impact of ORC1 on cilia. ORC1-deficiency confers defects anticipated to arise from impaired cilia function such as formation of oedema, kidney cysts, curved bodies and left–right asymmetry defects. We found ORC1 furthermore required for cilium formation in zebrafish and demonstrate that ciliopathy phenotypes in ORC1-depleted zebrafish could not be rescued by reconstitution with ORC1 bearing a genetic variant previously identified in MGS patients. Loss-of-function of Orc4 and Orc6, respectively, conferred similar ciliopathy phenotypes and cilium shortening in zebrafish, suggesting that several, if not all, components of the ORC regulate ciliogenesis downstream to or in addition to their canonical function in replication initiation. This study presents the first in vivo evidence of an influence of the MGS genes of the ORC family on cilia, and consolidates the possibility that cilia dysfunction could contribute to the clinical manifestation of ORC-deficient MGS.

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Acknowledgements

We thank Sandra Burczyk and Sophia Aicher for excellent fish care.

Authors’ contributions

LM, TCT, JG and CD performed experiments, LM, TCT, JG and MP analysed data, PAJ and MP conceived the study, designed the experiments and wrote the manuscript.

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Author notes

  1. These authors contributed equally: Lars D. Maerz, Teresa Casar Tena.

Affiliations

  1. Institute of Biochemistry and Molecular Biology, Ulm University, 89081, Ulm, Germany

    • Lars D. Maerz
    • , Teresa Casar Tena
    • , Julian Gerhards
    • , Cornelia Donow
    •  & Melanie Philipp
  2. Genome Damage and Stability Centre, University of Sussex, Brighton, BN1 9RQ, UK

    • Penelope A. Jeggo

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Financial support

Medical Research Council (G0000502; and G0500897, both to PAJ); Deutsche Forschungsgemeinschaft [DFG PH144/4; and DFG PH144/6 to MP]; Boehringer Ingelheim Ulm University BioCenter (to MP). TCT and LDM were fellows of the International Graduate School in Molecular Medicine at Ulm University.

Conflict of interest

The authors declare that they have no conflict of interest.

Corresponding author

Correspondence to Melanie Philipp.

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DOI

https://doi.org/10.1038/s41431-019-0338-0