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Asterless is a scaffold for the onset of centriole assembly

Nature volume 467, pages 714–718 (2010)Cite this article

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Abstract

Centrioles are found in the centrosome core and, as basal bodies, at the base of cilia and flagella. Centriole assembly and duplication is controlled by Polo-like-kinase 4 (Plk4): these processes fail if Plk4 is downregulated and are promoted by Plk4 overexpression1,2,3,4,5,6,7. Here we show that the centriolar protein Asterless (Asl; human orthologue CEP152) provides a conserved molecular platform, the amino terminus of which interacts with the cryptic Polo box of Plk4 whereas the carboxy terminus interacts with the centriolar protein Sas-4 (CPAP in humans). Drosophila Asl and human CEP152 are required for the centrosomal loading of Plk4 in Drosophila and CPAP in human cells, respectively. Depletion of Asl or CEP152 caused failure of centrosome duplication; their overexpression led to de novo centriole formation in Drosophila eggs, duplication of free centrosomes in Drosophila embryos, and centrosome amplification in cultured Drosophila and human cells. Overexpression of a Plk4-binding-deficient mutant of Asl prevented centriole duplication in cultured cells and embryos. However, this mutant protein was able to promote microtubule organizing centre (MTOC) formation in both embryos and oocytes. Such MTOCs had pericentriolar material and the centriolar protein Sas-4, but no centrioles at their core. Formation of such acentriolar MTOCs could be phenocopied by overexpression of Sas-4 in oocytes or embryos. Our findings identify independent functions for Asl as a scaffold for Plk4 and Sas-4 that facilitates self-assembly and duplication of the centriole and organization of pericentriolar material.

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Figure 1: Asl recruits Plk4 to centrosomes for centriole duplication.
Figure 2: Asl overexpression causes de novo formation and amplification of centrioles.
Figure 3: Overexpression of Asl-M1 mutant or the Asl interactor Sas-4 leads to loss of centrioles.
Figure 4: CEP152 interacts with human PLK4 and CPAP and controls centrosome duplication in human cells.

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Acknowledgements

We thank E. Wegel for technical help. We thank P. Coelho, G. Mao, P. Gönczy, T. Megraw and J. Raff for antibodies. We thank Cancer Research UK for a Programme Grant to D.M.G. and a studentship to Q.D.Y. K.W. was a visiting scholar of the Winston Churchill Foundation of the United States. Fundação Calouste Gulbenkian, Fundação para a Ciência e Tecnologia (FCT) provided support to M.B.-D. and scholarships to I.C.-F. and A.R.-M. The Royal Society provided an International Joint Project Grant for collaboration between M.B.-D. and D.M.G.

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Author notes

  1. Kipp Weiskopf

    Present address: Present Address: Stanford University School of Medicine, Stanford, California 94305, USA.,

  2. Nikola S. Dzhindzhev and Quan D. Yu: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Genetics, Cancer Research UK Cell Cycle Genetics Group, University of Cambridge, Downing Street, Cambridge, CB2 3EH, UK

    Nikola S. Dzhindzhev, Quan D. Yu, Kipp Weiskopf, George Tzolovsky, Ines Cunha-Ferreira, Ana Rodrigues-Martins & David M. Glover

  2. Instituto Gulbenkian de Ciencia, Oeiras, P-2780-156, Portugal

    Ines Cunha-Ferreira, Ana Rodrigues-Martins & Monica Bettencourt-Dias

  3. Dipartimento di Biologia Evolutiva, Università di Siena, via Aldo Moro, 2-53100 Siena, Italy,

    Maria Riparbelli & Giuliano Callaini

Authors
  1. Nikola S. Dzhindzhev
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Contributions

N.S.D. undertook interaction assays, mutagenesis and Drosophila cell culture work; Q.D.Y. worked on Drosophila embryos/eggs and de novo centriole formation; K.W. and I.C.-F. performed PrA–Plk4/Asl purifications; K.W. studied Asl depletion/overexpression in Drosophila cell culture. G.T. performed the human cell culture experiments. A.R.-M. and M.B.-D. overexpressed Sas-4 in embryos/eggs. M.R. and G.C. performed EM. N.S.D. and D.M.G. planned experiments and wrote the paper that was discussed by all authors.

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Correspondence to Nikola S. Dzhindzhev or David M. Glover.

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The authors declare no competing financial interests.

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Dzhindzhev, N., Yu, Q., Weiskopf, K. et al. Asterless is a scaffold for the onset of centriole assembly. Nature 467, 714–718 (2010). https://doi.org/10.1038/nature09445

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