Article | Published:

The Cep63 paralogue Deup1 enables massive de novo centriole biogenesis for vertebrate multiciliogenesis

Nature Cell Biology volume 15, pages 14341444 (2013) | Download Citation

Abstract

Dense multicilia in higher vertebrates are important for luminal flow and the removal of thick mucus. To generate hundreds of basal bodies for multiciliogenesis, specialized terminally differentiated epithelial cells undergo massive centriole amplification. In proliferating cells, however, centriole duplication occurs only once per cell cycle. How cells ensure proper regulation of centriole biogenesis in different contexts is poorly understood. We report that the centriole amplification is controlled by two duplicated genes, Cep63 and Deup1. Cep63 regulates mother-centriole-dependent centriole duplication. Deup1 governs deuterosome assembly to mediate large-scale de novo centriole biogenesis. Similarly to Cep63, Deup1 binds to Cep152 and then recruits Plk4 to activate centriole biogenesis. Phylogenetic analyses suggest that Deup1 diverged from Cep63 in a certain ancestor of lobe-finned fishes during vertebrate evolution and was subsequently adopted by tetrapods. Thus, the Cep63 gene duplication has enabled mother-centriole-independent assembly of the centriole duplication machinery to satisfy different requirements for centriole number.

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References

  1. 1.

    , & When cilia go bad: cilia defects and ciliopathies. Nat. Rev. Mol. Cell. Biol. 8, 880–893 (2007).

  2. 2.

    & Centrioles, centrosomes, and cilia in health and disease. Cell 139, 663–678 (2009).

  3. 3.

    & Basal body/centriole assembly and continuity. Curr. Opin. Cell Biol. 15, 96–104 (2003).

  4. 4.

    , & From zero to many: control of centriole number in development and disease. Traffic 10, 482–498 (2009).

  5. 5.

    & The centrosome cycle: centriole biogenesis, duplication and inherent asymmetries. Nat. Cell Biol. 13, 1154–1160 (2011).

  6. 6.

    , , , & Development and functions of the cytoskeleton during ciliogenesis in metazoa. Biol. Cell 63, 195–208 (1988).

  7. 7.

    & The formation of basal bodies (centrioles) in the Rhesus monkey oviduct. J. Cell Biol. 50, 10–34 (1971).

  8. 8.

    Reconstructions of centriole formation and ciliogenesis in mammalian lungs. J. Cell Sci. 3, 207–230 (1968).

  9. 9.

    Towards a molecular architecture of centriole assembly. Nat. Rev. Mol. Cell Biol. 13, 425–435 (2012).

  10. 10.

    & Building the centriole. Curr. Biol. 20, R816–825 (2010).

  11. 11.

    , , , & Cep152 interacts with Plk4 and is required for centriole duplication. J. Cell Biol. 191, 721–729 (2010).

  12. 12.

    et al. Cep152 acts as a scaffold for recruitment of Plk4 and CPAP to the centrosome. J. Cell Biol. 191, 731–739 (2010).

  13. 13.

    et al. Asterless is a scaffold for the onset of centriole assembly. Nature 467, 714–718 (2010).

  14. 14.

    et al. A primary microcephaly protein complex forms a ring around parental centrioles. Nat. Genet. 43, 1147–1153 (2011).

  15. 15.

    , , & Subdiffraction imaging of centrosomes reveals higher-order organizational features of pericentriolar material. Nat. Cell Biol. 14, 1148–1158 (2012).

  16. 16.

    et al. Subdiffraction-resolution fluorescence microscopy reveals a domain of the centrosome critical for pericentriolar material organization. Nat. Cell Biol. 14, 1159–1168 (2012).

  17. 17.

    , , & 3D-structured illumination microscopy provides novel insight into architecture of human centrosomes. Biol. Open 1, 965–976 (2012).

  18. 18.

    et al. Plk4-induced centriole biogenesis in human cells. Dev. Cell 13, 190–202 (2007).

  19. 19.

    et al. The human microcephaly protein STIL interacts with CPAP and is required for procentriole formation. EMBO J. 30, 4790–4804 (2011).

  20. 20.

    , , , & CPAP is a cell-cycle regulated protein that controls centriole length. Nat. Cell Biol. 11, 825–831 (2009).

  21. 21.

    et al. Overly long centrioles and defective cell division on excess of the SAS-4-related protein CPAP. Curr. Biol. 19, 1012–1018 (2009).

  22. 22.

    et al. Control of centriole length by CPAP and CP110. Curr. Biol. 19, 1005–1011 (2009).

  23. 23.

    , , & Cell-cycle-regulated expression of STIL controls centriole number in human cells. J. Cell Sci. 125, 1342–1352 (2012).

  24. 24.

    et al. STIL is required for centriole duplication in human cells. J. Cell Sci. 125, 1353–1362 (2012).

  25. 25.

    et al. Regulated HsSAS-6 levels ensure formation of a single procentriole per centriole during the centrosome duplication cycle. Dev. Cell 13, 203–213 (2007).

  26. 26.

    Centriole morphogenesis in developing ciliated epithelium of the mouse oviduct. J. Cell Biol. 51, 286–302 (1971).

  27. 27.

    , , & Growth and differentiation of mouse tracheal epithelial cells: selection of a proliferative population. Am. J. Physiol. Lung. Cell Mol. Physiol. 283, L1315–1321 (2002).

  28. 28.

    & Molecular characterization of centriole assembly in ciliated epithelial cells. J. Cell Biol. 178, 31–42 (2007).

  29. 29.

    et al. miR-129-3p controls cilia assembly by regulating CP110 and actin dynamics. Nat. Cell Biol. 14, 697–706 (2012).

  30. 30.

    & Monoclonal antibodies specific for an acetylated form of alpha-tubulin recognize the antigen in cilia and flagella from a variety of organisms. J. Cell Biol. 101, 2085–2094 (1985).

  31. 31.

    & Building a centriole. Curr. Opin. Cell Biol. 25, 72–77 (2013).

  32. 32.

    , , & The Polo kinase Plk4 functions in centriole duplication. Nat. Cell Biol. 7, 1140–1146 (2005).

  33. 33.

    et al. Loss of C. elegans BBS-7 and BBS-8 protein function results in cilia defects and compromised intraflagellar transport. Genes. Dev. 18, 1630–1642 (2004).

  34. 34.

    & The intraflagellar transport protein IFT57 is required for cilia maintenance and regulates IFT-particle-kinesin-II dissociation in vertebrate photoreceptors. J. Cell Sci. 121, 1907–1915 (2008).

  35. 35.

    , , & Outer dense fibre 2 is a widespread centrosome scaffold component preferentially associated with mother centrioles: its identification from isolated centrosomes. Mol. Biol. Cell 12, 1687–1697 (2001).

  36. 36.

    & The complete DNA sequence of the mitochondrial genome of a ‘living fossil,’ the coelacanth (Latimeria chalumnae). Genetics 146, 995–1010 (1997).

  37. 37.

    et al. The African coelacanth genome provides insights into tetrapod evolution. Nature 496, 311–316 (2013).

  38. 38.

    & Evolutionary crossroads in developmental biology: amphioxus. Development 138, 4819–4830 (2011).

  39. 39.

    , & Seeing chordate evolution through the Ciona genome sequence. Genome. Biol. 4, 208 (2003).

  40. 40.

    et al. Identification of novel ciliogenesis factors using a new in vivo model for mucociliary epithelial development. Dev. Biol. 312, 115–130 (2007).

  41. 41.

    & Nephrocystins and MKS proteins interact with IFT particle and facilitate transport of selected ciliary cargos. EMBO J. 30, 2532–2544 (2011).

  42. 42.

    , , , & Nudel is crucial for the WAVE complex assembly in vivo by selectively promoting subcomplex stability and formation through direct interactions. Cell Res. 22, 1270–1284 (2012).

  43. 43.

    , & Posttranslational quality control: folding, refolding, and degrading proteins. Science 286, 1888–1893 (1999).

  44. 44.

    et al. The m-subunit of murine translation initiation factor eIF3 maintains the integrity of the eIF3 complex and is required for embryonic development, homeostasis, and organ size control. J. Biol. Chem. 288, 30087–30093 (2013).

  45. 45.

    & Basal body assembly in ciliates: the power of numbers. Traffic 10, 461–471 (2009).

  46. 46.

    , , , & Centrosome loss in the evolution of planarians. Science 335, 461–463 (2012).

  47. 47.

    & Gene and genome duplications in vertebrates: the one-to-four (-to-eight in fish) rule and the evolution of novel gene functions. Curr. Opin. Cell Biol. 11, 699–704 (1999).

  48. 48.

    et al. Cilia-driven fluid flow in the zebrafish pronephros, brain and Kupffer’s vesicle is required for normal organogenesis. Development 132, 1907–1921 (2005).

  49. 49.

    The role of epidermal cilia in development of the Australian lungfish, Neoceratodus forsteri (Osteichthyes: Dipnoi). J. Morphol. 228, 203–221 (1996).

  50. 50.

    , & A guide to super-resolution fluorescence microscopy. J. Cell Biol. 190, 165–175 (2010).

  51. 51.

    , , , & The microtubule plus end-binding protein EB1 is involved in Sertoli cell plasticity in testicular seminiferous tubules. Exp. Cell Res. 314, 213–226 (2008).

  52. 52.

    , , & Notch signalling controls the differentiation of transporting epithelia and multiciliated cells in the zebrafish pronephros. Development 134, 1111–1122 (2007).

  53. 53.

    & Natural mating and tadpole husbandry in the western clawed frog Xenopus tropicalis. Cold. Spring Harbor. Protocols. 2009 pdb prot5292 (2009).

  54. 54.

    , , & Regulation of cell-matrix adhesion dynamics and Rac-1 by integrin linked kinase. FASEB J. 20, 1489–1491 (2006).

  55. 55.

    , & Assembly of the FtsZ ring at the central division site in the absence of the chromosome. Mol. Microbiol. 29, 491–503 (1998).

  56. 56.

    et al. Nudel and FAK as antagonizing strength modulators of nascent adhesions through paxillin. PLoS Biol. 7 e1000116 (2009).

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Acknowledgements

The authors thank Y. Zheng, Y. Shen and J. Ten for helpful suggestions; E. Nigg for Cep152 antibody; S. Li and Z. Li for technical support on 3D-SIM and H. Dai, T. Zhang, F. Qin and X. Zhu for technical assistance. This work was supported by the Chinese Academy of Sciences (XDA01010107), the National Basic Research Program of China (2012CB945003 and 2010CB912102), the National Science Foundation of China (31330045, 31271427 and 91129000) and Shanghai Municipal Science and Technology Commission (12JC1409900). X.Y. acknowledges the support of the SA-SIBS Scholarship Program.

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Author notes

    • Xiumin Yan
    •  & Xueliang Zhu

    These authors contributed equally to this work

Affiliations

  1. State Key Laboratory of Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China

    • Huijie Zhao
    • , Lei Zhu
    • , Yunlu Zhu
    • , Jingli Cao
    • , Shanshan Li
    • , Qiongping Huang
    • , Xiumin Yan
    •  & Xueliang Zhu
  2. Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Beijing 100101, China

    • Tao Xu
  3. College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang 310058, China

    • Xiao Huang

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Contributions

X.Y. and X.Z. conceived and directed the project. H.Z. carried out most experimental work. L.Z. carried out the frog and transmission EM experiments. Y.Z did the experiments in bacteria and drew all illustrative models. J.C. established the culture and infection conditions for MTECs. S.L. found that CEP152 RNAi downregulated CEP63. Q.H. generated the homemade antibodies. T.X. and X.H. provided the 3D-SIM and frog systems. X.Z., X.Y. and H.Z. designed experiments, interpreted data, and wrote the paper.

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The authors declare no competing financial interests.

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Correspondence to Xiumin Yan or Xueliang Zhu.

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Videos

  1. 1.

    3D model of a part of a cell in stage II immunostained for Deup1 (green), Cep152 (blue), and Centrin (red).

    The top and side views are represented in Fig. 4c.

  2. 2.

    3D model of a part of a cell in stage III immunostained for Deup1 (green), Cep152 (blue), and Centrin (red).

    The top and side views are represented in Fig. 4c.

  3. 3.

    3D model of a part of a cell in stage IV immunostained for Deup1 (green), Cep152 (blue), and Centrin (red).

    The top and side views are represented in Fig. 4c.

  4. 4.

    3D model of a part of a cell in stage V immunostained for Deup1 (green), Cep152 (blue), and Centrin (red).

    The top and side views are represented in Fig. 4c.

  5. 5.

    3D model of a part of a cell in stage VI immunostained for Deup1 (green), Cep152 (blue), and Centrin (red).

    The top and side views are represented in Fig. 4c.

  6. 6.

    EM images from four consecutive sections of a control (Ctrl-i) and a Deup1-knockdown (Dp-i1) MTEC at ALI d3.

    The thickness of each section is 70 nm. Deuterosomes (d), mother centrioles (m), and daughter procentrioles around mother (p) are numbered according to the sequence of appearance. The overlaid images are shown in Fig. 7a.

  7. 7.

    3D model for deuterosome-like structures formed by His-Deup1 (green) in E. coli.

    The protein expression was induced with 10 μM IPTG. See Fig. 7d for the original 3D-SIM image.

  8. 8.

    3D model for deuterosome-like structures formed by His-Deup1 (green) in E. coli.

    The protein expression was induced with 50 μM IPTG. See Fig. 7d for the original 3D-SIM image.

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DOI

https://doi.org/10.1038/ncb2880

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