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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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.).
The authors declare no competing financial interests.
Supplementary Note, Supplementary Figures 1–12, Supplementary Table 2 (PDF 3157 kb)
Spreadsheet of SILAC results (XLS 34 kb)
Primer sequences (XLSX 45 kb)
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)
Mitosis in CEP63KO DT40 cells expressing GFP-α-tubulin. Images were acquired at a rate of 3 minutes/frame. (MOV 1504 kb)
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)
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)
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)
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)
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)
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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