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SADB phosphorylation of γ-tubulin regulates centrosome duplication

Nature Cell Biology volume 11, pages 10811092 (2009) | Download Citation

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Abstract

Symmetrical cell division requires duplication of DNA and protein content to generate two daughter cells. Centrosomes also duplicate during cell division, but the mechanism controlling this process is incompletely understood. We describe an alternative splice form of SadB encoding a short SADB Ser/Thr kinase whose activity fluctuates during the cell cycle, localizes to centrosomes, and controls centrosome duplication. Reduction of endogenous SADB levels diminished centrosome numbers, whereas enhanced SADB expression induced centrosome amplification. SADB exerted this action through phosphorylation of γ-tubulin on Ser 131, as expression of a phosphomimetic Ser 131-to-Asp γ-tubulin mutant alone increased centrosome numbers, whereas non-phosphorylatable Ala 131-γ-tubulin impaired centrosome duplication. We propose that SADB kinase activity controls centrosome homeostasis by regulating phosphorylation of γ-tubulin.

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Acknowledgements

We thank York-Dieter Stierhof and Dagmar Ripper for their generous help with electron microscopy. We thank Y. Shi, M. van de Wetering, M. Marqués, M. Bornens and T. Strearns for reagents, J. Sanes and B. Lilley for SADB-deficient cells and comments, L. Kremer and L. Gómez for antibody production, C. Hernández for help in biochemistry assays, C. Mark and R. Hartong for editorial assistance. This work was supported by the Spanish Ministry of Science and Innovation (SAF200405955, 200763624 to ACC, CSD 2006-00023 to JMV), the Spanish Association against Cancer, the Spanish Ministry of Health (PI050964), the Royal Physiographic Society in Lund, the Ake Wibergs, Thelma Zoegas, OE och Edla Johanssons and U-MAS Cancer Research Found, and fellowships from Teggers Fond, EMBO and the Swedish Society for Medical Research.

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Affiliations

  1. Department of Immunology and Oncology, Centro Nacional de Biotecnología/CSIC, Darwin 3, Madrid E-28049, Spain.

    • María Alvarado-Kristensson
    • , Virginia Silió
    •  & Ana C. Carrera
  2. Center for Molecular Pathology, Dept. Laboratory Medicine, Lund University, Malmoe University Hospital, Sweden.

    • María Alvarado-Kristensson
  3. Department of Macromolecular Structures, Centro Nacional de Biotecnología/CSIC, Darwin 3, Madrid E-28049, Spain.

    • María Josefa Rodríguez
    •  & José M. Valpuesta

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Contributions

M.A.K. performed most experiments; M.J.R. and J.M.V. performed EM experiments; V.S. performed some experiments and A.C.C. directed the work, performed some assays and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to María Alvarado-Kristensson or Ana C. Carrera.

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DOI

https://doi.org/10.1038/ncb1921

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