Structural Basis for Mitotic Centrosome Assembly in Flies.
Haensele, Andreas FM
Cottee, Matthew A
Lea, Susan M
Raff, Jordan W
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Feng, Z., Caballe, A., Wainman, A., Johnson, S., Haensele, A. F., Cottee, M. A., Conduit, P., et al. (2017). Structural Basis for Mitotic Centrosome Assembly in Flies.. Cell, 169 (6), 1078-1089.e13. https://doi.org/10.1016/j.cell.2017.05.030
In flies, Centrosomin (Cnn) forms a phosphorylation-dependent scaffold that recruits proteins to the mitotic centrosome, but how Cnn assembles into a scaffold is unclear. We show that scaffold assembly requires conserved leucine zipper (LZ) and Cnn-motif 2 (CM2) domains that co-assemble into a 2:2 complex in vitro. We solve the crystal structure of the LZ:CM2 complex, revealing that both proteins form helical dimers that assemble into an unusual tetramer. A slightly longer version of the LZ can form micron-scale structures with CM2, whose assembly is stimulated by Plk1 phosphorylation in vitro. Mutating individual residues that perturb LZ:CM2 tetramer assembly perturbs the formation of these micron-scale assemblies in vitro and Cnn-scaffold assembly in vivo. Thus, Cnn molecules have an intrinsic ability to form large, LZ:CM2-interaction-dependent assemblies that are critical for mitotic centrosome assembly. These studies provide the first atomic insight into a molecular interaction required for mitotic centrosome assembly.
Centrosomin, Cnn, PCM, Plk1, centriole, centrosome, mitosis, Amino Acid Sequence, Animals, Centrosome, Drosophila Proteins, Drosophila melanogaster, Homeodomain Proteins, Mitosis, Models, Molecular, Phosphorylation, Protein Domains, Protein-Serine-Threonine Kinases, Sequence Alignment
Z.F. and A.F.M.H. were supported by Sir William Dunn School EPA PhD studentships and also a Clarendon Scholarship and a Santander Graduate Award to A.F.M.H; A.C., A.W., M.A.C., P.T.C., and J.W.R. were supported by a Wellcome Trust Senior Investigator Award (104575); S.J. and S.M.L. were supported by a Wellcome Trust Senior Investigator Award (100298); A.W. was also partially supported by a Wellcome Trust Strategic Award to the Micron Oxford Advanced Bioimaging Unit (107457).
WELLCOME TRUST (105653/Z/14/Z)
External DOI: https://doi.org/10.1016/j.cell.2017.05.030
This record's URL: https://www.repository.cam.ac.uk/handle/1810/266598
Attribution 4.0 International
Licence URL: http://creativecommons.org/licenses/by/4.0/