the role of γ-tubulin in centrosomal microtubule organizationγ-tubulin centrosomal微管组织的作用.pdfVIP

The Role of c-Tubulin in Centrosomal Microtubule Organization 1 2¤ 3 3 ¨ 4 Eileen O’Toole , Garrett Greenan , Karen I. Lange , Martin Srayko , Thomas Muller-Reichert * 1 Boulder Laboratory for 3-D Electron Microscopy of Cells, University of Colorado Boulder, Boulder, Colorado, United States of America, 2 Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany, 3 Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada, 4 Medical Theoretical Center, University of Technology Dresden, Dresden, Germany Abstract As part of a multi-subunit ring complex, c-tubulin has been shown to promote microtubule nucleation both in vitro and in vivo, and the structural properties of the complex suggest that it also seals the minus ends of the polymers with a conical cap. Cells depleted of c-tubulin, however, still display many microtubules that participate in mitotic spindle assembly, suggesting that c-tubulin is not absolutely required for microtubule nucleation in vivo, and raising questions about the function of the minus end cap. Here, we assessed the role of c-tubulin in centrosomal microtubule organisation using three- dimensional reconstructions of c-tubulin-depleted C. elegans embryos. We found that microtubule minus-end capping and the PCM component SPD-5 are both essential for the proper placement of microtubules in the centrosome. Our results further suggest that c-tubulin and SPD-5 limit microtubule polymerization within the centrosome core, and we propose a model for how abnormal microtubule organization at the centrosome could indirectly affect centriole structure and daughter centriole replication