A primary microcephaly protein complex forms a ring around parental centrioles


Autosomal recessive primary microcephaly (MCPH) is characterized by a substantial reduction in prenatal human brain growth without alteration of the cerebral architecture and is caused by biallelic mutations in genes coding for a subset of centrosomal proteins1,2,3,4,5,6,7,8,9,10. Although at least three of these proteins have been implicated in centrosome duplication11, the nature of the centrosome dysfunction that underlies the neurodevelopmental defect in MCPH is unclear. Here we report a homozygous MCPH-causing mutation in human CEP63. CEP63 forms a complex with another MCPH protein, CEP152, a conserved centrosome duplication factor12,13,14,15. Together, these two proteins are essential for maintaining normal centrosome numbers in cells. Using super-resolution microscopy, we found that CEP63 and CEP152 co-localize in a discrete ring around the proximal end of the parental centriole, a pattern specifically disrupted in CEP63-deficient cells derived from patients with MCPH. This work suggests that the CEP152-CEP63 ring-like structure ensures normal neurodevelopment and that its impairment particularly affects human cerebral cortex growth.

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Figure 1: Identification of an MCPH-causing mutation in CEP63.
Figure 2: Disruption of the centrosomal gene CEP63 in vertebrate cells.
Figure 3: CEP63 is required for maintaining normal centrosome numbers.
Figure 4: CEP63 forms a protein complex with CEP152.
Figure 5: CEP63-dependent centrosomal accumulation of CEP152 maintains normal centrosome numbers.
Figure 6: CEP63 and CEP152 form a ring around parental centrioles, a structure disrupted in CEP63−/− cells from affected individuals.

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  1. 1

    Woods, C.G., Bond, J. & Enard, W. Autosomal recessive primary microcephaly (MCPH): a review of clinical, molecular and evolutionary findings. Am. J. Hum. Genet. 76, 717–728 (2005).

    CAS  Article  Google Scholar 

  2. 2

    Jackson, A.P. et al. Identification of microcephalin, a protein implicated in determining the size of the human brain. Am. J. Hum. Genet. 71, 136–142 (2002).

    CAS  Article  Google Scholar 

  3. 3

    Bond, J. et al. ASPM is a major determinant of cerebral cortical size. Nat. Genet. 32, 316–320 (2002).

    CAS  Article  Google Scholar 

  4. 4

    Bond, J. et al. A centrosomal mechanism involving CDK5RAP2 and CENPJ controls brain size. Nat. Genet. 37, 353–355 (2005).

    CAS  Article  Google Scholar 

  5. 5

    Kumar, A., Girimaji, S.C., Duvvari, M.R. & Blanton, S.H. Mutations in STIL, encoding a pericentriolar and centrosomal protein, cause primary microcephaly. Am. J. Hum. Genet. 84, 286–290 (2009).

    CAS  Article  Google Scholar 

  6. 6

    Nicholas, A.K. et al. WDR62 is associated with the spindle pole and is mutated in human microcephaly. Nat. Genet. 42, 1010–1014 (2010).

    CAS  Article  Google Scholar 

  7. 7

    Yu, T.W. et al. Mutations in WDR62, encoding a centrosome-associated protein, cause microcephaly with simplified gyri and abnormal cortical architecture. Nat. Genet. 42, 1015–1020 (2010).

    CAS  Article  Google Scholar 

  8. 8

    Bilgüvar, K. et al. Whole-exome sequencing identifies recessive WDR62 mutations in severe brain malformations. Nature 467, 207–210 (2010).

    Article  Google Scholar 

  9. 9

    Guernsey, D.L. et al. Mutations in centrosomal protein CEP152 in primary microcephaly families linked to MCPH4. Am. J. Hum. Genet. 87, 40–51 (2010).

    CAS  Article  Google Scholar 

  10. 10

    Thornton, G.K. & Woods, C.G. Primary microcephaly: do all roads lead to Rome? Trends Genet. 25, 501–510 (2009).

    CAS  Article  Google Scholar 

  11. 11

    Bettencourt-Dias, M., Hildebrandt, F., Pellman, D., Woods, G. & Godinho, S.A. Centrosomes and cilia in human disease. Trends Genet. 27, 307–315 (2011).

    CAS  Article  Google Scholar 

  12. 12

    Blachon, S. et al. Drosophila asterless and vertebrate Cep152 are orthologs essential for centriole duplication. Genetics 180, 2081–2094 (2008).

    CAS  Article  Google Scholar 

  13. 13

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

    CAS  Article  Google Scholar 

  14. 14

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

    CAS  Article  Google Scholar 

  15. 15

    Hatch, E.M., Kulukian, A., Holland, A.J., Cleveland, D.W. & Stearns, T. Cep152 interacts with Plk4 and is required for centriole duplication. J. Cell Biol. 191, 721–729 (2010).

    CAS  Article  Google Scholar 

  16. 16

    Roberts, E. et al. Autosomal recessive primary microcephaly: an analysis of locus heterogeneity and phenotypic variation. J. Med. Genet. 39, 718–721 (2002).

    CAS  Article  Google Scholar 

  17. 17

    Andersen, J.S. et al. Proteomic characterization of the human centrosome by protein correlation profiling. Nature 426, 570–574 (2003).

    CAS  Article  Google Scholar 

  18. 18

    Evans, P.D. et al. Microcephalin, a gene regulating brain size, continues to evolve adaptively in humans. Science 309, 1717–1720 (2005).

    CAS  Article  Google Scholar 

  19. 19

    Mekel-Bobrov, N. et al. Ongoing adaptive evolution of ASPM, a brain size determinant in Homo sapiens. Science 309, 1720–1722 (2005).

    CAS  Article  Google Scholar 

  20. 20

    Evans, P.D., Vallender, E.J. & Lahn, B.T. Molecular evolution of the brain size regulator genes CDK5RAP2 and CENPJ. Gene 375, 75–79 (2006).

    CAS  Article  Google Scholar 

  21. 21

    Tang, B.L. Molecular genetic determinants of human brain size. Biochem. Biophys. Res. Commun. 345, 911–916 (2006).

    CAS  Article  Google Scholar 

  22. 22

    Smith, E. et al. An ATM- and ATR-dependent checkpoint inactivates spindle assembly by targeting CEP63. Nat. Cell Biol. 11, 278–285 (2009).

    CAS  Article  Google Scholar 

  23. 23

    Löffler, H. et al. Cep63 recruits Cdk1 to the centrosome: implications for regulation of mitotic entry, centrosome amplification, and genome maintenance. Cancer Res. 71, 2129–2139 (2011).

    Article  Google Scholar 

  24. 24

    Bürckstümmer, T. et al. An efficient tandem affinity purification procedure for interaction proteomics in mammalian cells. Nat. Methods 3, 1013–1019 (2006).

    Article  Google Scholar 

  25. 25

    Bettencourt-Dias, M. & Glover, D.M. Centrosome biogenesis and function: centrosomics brings new understanding. Nat. Rev. Mol. Cell Biol. 8, 451–463 (2007).

    CAS  Article  Google Scholar 

  26. 26

    Barr, A.R., Kilmartin, J.V. & Gergely, F. CDK5RAP2 functions in centrosome to spindle pole attachment and DNA damage response. J. Cell Biol. 189, 23–39 (2010).

    CAS  Article  Google Scholar 

  27. 27

    Mayer, T.U. et al. Small molecule inhibitor of mitotic spindle bipolarity identified in a phenotype-based screen. Science 286, 971–974 (1999).

    CAS  Article  Google Scholar 

  28. 28

    Tsou, M.-F.B. & Stearns, T. Mechanism limiting centrosome duplication to once per cell cycle. Nature 442, 947–951 (2006).

    CAS  Article  Google Scholar 

  29. 29

    Nigg, E.A. Centrosome duplication: of rules and licenses. Trends Cell Biol. 17, 215–221 (2007).

    CAS  Article  Google Scholar 

  30. 30

    Loncarek, J., Hergert, P. & Khodjakov, A. Centriole reduplication during prolonged interphase requires procentriole maturation governed by Plk1. Curr. Biol. 20, 1277–1282 (2010).

    CAS  Article  Google Scholar 

  31. 31

    Wang, W.J., Soni, R.K., Uryu, K. & Bryan Tsou, M.F. The conversion of centrioles to centrosomes: essential coupling of duplication with segregation. J. Cell Biol. 193, 727–739 (2011).

    CAS  Article  Google Scholar 

  32. 32

    Ong, S.E. et al. Stable isotope labeling by amino acids in cell culture, SILAC, as a simple and accurate approach to expression proteomics. Mol. Cell. Proteomics 1, 376–386 (2002).

    CAS  Article  Google Scholar 

  33. 33

    Hutchins, J.R. et al. Systematic analysis of human protein complexes identifies chromosome segregation proteins. Science 328, 593–599 (2010).

    CAS  Article  Google Scholar 

  34. 34

    Bonaccorsi, S., Giansanti, M.G. & Gatti, M. Spindle self-organization and cytokinesis during male meiosis in asterless mutants of Drosophila melanogaster. J. Cell Biol. 142, 751–761 (1998).

    CAS  Article  Google Scholar 

  35. 35

    Varmark, H. et al. Asterless is a centriolar protein required for centrosome function and embryo development in Drosophila. Curr. Biol. 17, 1735–1745 (2007).

    CAS  Article  Google Scholar 

  36. 36

    Kalay, E. et al. CEP152 is a genome maintenance protein disrupted in Seckel syndrome. Nat. Genet. 43, 23–26 (2011).

    CAS  Article  Google Scholar 

  37. 37

    Gillingham, A.K. & Munro, S. The PACT domain, a conserved centrosomal targeting motif in the coiled-coil proteins AKAP450 and pericentrin. EMBO Rep. 1, 524–529 (2000).

    CAS  Article  Google Scholar 

  38. 38

    Leidel, S., Delattre, M., Cerutti, L., Baumer, K. & Gonczy, P. SAS-6 defines a protein family required for centrosome duplication in C. elegans and in human cells. Nat. Cell Biol. 7, 115–125 (2005).

    CAS  Article  Google Scholar 

  39. 39

    Tsou, M.F. et al. Polo kinase and separase regulate the mitotic licensing of centriole duplication in human cells. Dev. Cell 17, 344–354 (2009).

    CAS  Article  Google Scholar 

  40. 40

    Lénárt, P. et al. The small-molecule inhibitor BI 2536 reveals novel insights into mitotic roles of polo-like kinase 1. Curr. Biol. 17, 304–315 (2007).

    Article  Google Scholar 

  41. 41

    Bettencourt-Dias, M. et al. SAK/PLK4 is required for centriole duplication and flagella development. Curr. Biol. 15, 2199–2207 (2005).

    CAS  Article  Google Scholar 

  42. 42

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

    CAS  Article  Google Scholar 

  43. 43

    Puklowski, A. et al. The SCF-Fbxw5 E3-ubiquitin ligase is regulated by Plk4 and targets HsSAS-6 to control centrosome duplication. Nat. Cell Biol. 13, 1004–1009 (2011).

    CAS  Article  Google Scholar 

  44. 44

    Rogers, G.C., Rusan, N.M., Roberts, D.M., Peifer, M. & Rogers, S.L. The SCF Slimb ubiquitin ligase regulates Plk4/Sak levels to block centriole reduplication. J. Cell Biol. 184, 225–239 (2009).

    CAS  Article  Google Scholar 

  45. 45

    Calegari, F., Haubensak, W., Haffner, C. & Huttner, W.B. Selective lengthening of the cell cycle in the neurogenic subpopulation of neural progenitor cells during mouse brain development. J. Neurosci. 25, 6533–6538 (2005).

    CAS  Article  Google Scholar 

  46. 46

    Keryer, G. et al. Dissociating the centrosomal matrix protein AKAP450 from centrioles impairs centriole duplication and cell cycle progression. Mol. Biol. Cell 14, 2436–2446 (2003).

    CAS  Article  Google Scholar 

  47. 47

    Stevens, N.R., Roque, H. & Raff, J.W. DSas-6 and Ana2 coassemble into tubules to promote centriole duplication and engagement. Dev. Cell 19, 913–919 (2010).

    CAS  Article  Google Scholar 

  48. 48

    Barrera, J.A. et al. CDK5RAP2 regulates centriole engagement and cohesion in mice. Dev. Cell 18, 913–926 (2010).

    CAS  Article  Google Scholar 

  49. 49

    Lizarraga, S.B. et al. Cdk5rap2 regulates centrosome function and chromosome segregation in neuronal progenitors. Development 137, 1907–1917 (2010).

    CAS  Article  Google Scholar 

  50. 50

    Wang, X. et al. Asymmetric centrosome inheritance maintains neural progenitors in the neocortex. Nature 461, 947–955 (2009).

    CAS  Article  Google Scholar 

  51. 51

    Darlington, G.J. Lymphocyte isolation and culture. Cold Spring Harb. Protoc. doi:10.1101/pdb.prot4480 (2006).

  52. 52

    Lewis, A.E. et al. Identification of nuclear phosphatidylinositol 4,5-bisphosphate-interacting proteins by neomycin extraction. Mol. Cell. Proteomics 10, M110.003376 (2011).

    Article  Google Scholar 

  53. 53

    Finch, A.J. et al. Uncoupling of GTP hydrolysis from eIF6 release on the ribosome causes Shwachman-Diamond syndrome. Genes Dev. 25, 917–929 (2011).

    CAS  Article  Google Scholar 

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We thank S. Munro, J. Pines and R. Rios for constructs and reagents; J. Cox, H. Ebrahimi and G. Thornton for their contribution; P. Lakshminarasimhan, K.J. Patel and R. Rios for critical reading of the manuscript; and M. Bornens and the Gergely lab for helpful suggestions. We also thank the families for their participation. We are grateful for the support provided by the microscopy and proteomics core facilities at the Cambridge Research Institute. This work was made possible by funding from the Wellcome Trust (to A.K.N., C.G.W., M.K. and O.P.C.), from Cancer Research UK (to A.R.B., C.D., F.G., J.-H.S. and S.R.) and a Royal Society University Research Fellowship (to F.G.).

Author information




J.-H.S. performed most of the experiments presented in the manuscript. A.R.B. generated affected cell lines and performed the initial cell biology analysis. A.K.N. performed molecular genetic mapping and gene mutation identification. O.P.C. performed gene expression analysis in affected cells. M.K. carried out embryonic brain immunohistochemistry. A.S. and S.R. provided support with the super-resolution microscopy. C.D. helped with generation and analysis of proteomic data. C.G.W. performed subject ascertainment, clinical studies and the gene-identification strategy. C.G.W. and F.G. designed the study and wrote the paper, with comments from all authors.

Corresponding authors

Correspondence to C Geoffrey Woods or Fanni Gergely.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Note, Supplementary Figures 1–12, Supplementary Table 2 (PDF 3157 kb)

Supplementary Table 1

Spreadsheet of SILAC results (XLS 34 kb)

Supplementary Table 3

Primer sequences (XLSX 45 kb)

Supplementary Video 1

Mitosis in wild-type DT40 cells expressing GFP-α-tubulin. Images were acquired at a rate of 3 minutes/frame. Note that software failed to assign certain frames with correct timestamps. (MOV 755 kb)

Supplementary Video 2

Mitosis in CEP63KO DT40 cells expressing GFP-α-tubulin. Images were acquired at a rate of 3 minutes/frame. (MOV 1504 kb)

Supplementary Video 3

Mitosis in wild-type DT40 cells expressing GFP-PACT and Ruby-Histone H2B. Images were acquired at a rate of 6 minutes/frame. GFP is green, ruby is red. (MOV 595 kb)

Supplementary Video 4

Mitosis in wild-type DT40 cells expressing GFP-PACT and Ruby-Histone H2B. Images were acquired at a rate of 6 minutes/frame. Note that one centrosome is weaker. GFP is green, ruby is red. (MOV 986 kb)

Supplementary Video 5

Mitosis in CEP63KO DT40 cells expressing GFP-PACT and Ruby-Histone H2B. Images were acquired at a rate of 6 minutes/frame. GFP is green, ruby is red. (MOV 753 kb)

Supplementary Video 6

Mitosis in CEP63KO DT40 cells expressing GFP-PACT and Ruby-Histone H2B. Images were acquired at a rate of 6 minutes/frame. GFP is green, ruby is red. (MOV 2089 kb)

Supplementary Video 7

Mitosis in CEP63KO DT40 cells expressing GFP-PACT and Ruby-Histone H2B. Images were acquired at a rate of 6 minutes/frame. GFP is green, ruby is red. (MOV 1186 kb)

Supplementary Video 8

Mitosis in CEP63KO DT40 cells expressing GFP-PACT and Ruby-Histone H2B. Images were acquired at a rate of 6 minutes/frame. GFP is green, ruby is red. (MOV 1415 kb)

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Sir, J., Barr, A., Nicholas, A. et al. A primary microcephaly protein complex forms a ring around parental centrioles. Nat Genet 43, 1147–1153 (2011). https://doi.org/10.1038/ng.971

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