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Reconstitution of the recombinant human γ-tubulin ring complex |
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| Authors: | Martin Würtz Anna Bhler Annett Neuner Erik Zupa Lukas Rohland Peng Liu Bram J A Vermeulen Stefan Pfeffer Sebastian Eustermann Elmar Schiebel |
| Institution: | 1. Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany ; 2. Centre for Organismal Studies Universität Heidelberg (COS), Im Neuenheimer Feld 230, D-69120 Heidelberg, Germany ; 3. European Molecular Biology Laboratory (EMBL), Heidelberg Meyerhofstraße 1, 69117 Heidelberg, Germany |
| Abstract: | Cryo-electron microscopy recently resolved the structure of the vertebrate γ-tubulin ring complex (γ-TuRC) purified from Xenopus laevis egg extract and human cells to near-atomic resolution. These studies clarified the arrangement and stoichiometry of γ-TuRC components and revealed that one molecule of actin and the small protein MZT1 are embedded into the complex. Based on this structural census of γ-TuRC core components, we developed a recombinant expression system for the reconstitution and purification of human γ-TuRC from insect cells. The recombinant γ-TuRC recapitulates the structure of purified native γ-TuRC and has similar functional properties in terms of microtubule nucleation and minus end capping. This recombinant system is a central step towards deciphering the activation mechanisms of the γ-TuRC and the function of individual γ-TuRC core components. |
| Keywords: | γ -tubulin microtubules γ -TuRC |