1. Structural Biology and Molecular Biophysics
  2. Cell Biology

The homo-oligomerisation of both Sas-6 and Ana2 is required for efficient centriole assembly in flies

  1. Matthew A Cottee
  2. Nadine Muschalik
  3. Steven Johnson
  4. Joanna Leveson
  5. Jordan W Raff  Is a corresponding author
  6. Susan M Lea
  1. University of Oxford, United Kingdom
  2. MRC-Laboratory of Molecular Biology, United Kingdom
https://doi.org/10.7554/eLife.07236
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  1. Matthew A Cottee
  2. Nadine Muschalik
  3. Steven Johnson
  4. Joanna Leveson
  5. Jordan W Raff
  6. Susan M Lea
(2015)
The homo-oligomerisation of both Sas-6 and Ana2 is required for efficient centriole assembly in flies
eLife 4:e07236.
https://doi.org/10.7554/eLife.07236
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Abstract

Sas-6 and Ana2/STIL proteins are required for centriole duplication and the homo-oligomerisation properties of Sas-6 help establish the nine-fold symmetry of the central cartwheel that initiates centriole assembly. Ana2/STIL proteins are poorly conserved, but they all contain a predicted Central Coiled-Coil Domain (CCCD). Here we show that the Drosophila Ana2 CCCD forms a tetramer, and we solve its structure to 0.8 Å, revealing that it adopts an unusual parallel-coil topology. We also solve the structure of the Drosophila Sas-6 N-terminal domain to 2.9 Å revealing that it forms higher-order oligomers through canonical interactions. Point mutations that perturb Sas-6 or Ana2 homo-oligomerisation in vitro strongly perturb centriole assembly in vivo. Thus, efficient centriole duplication in flies requires the homo-oligomerisation of both Sas-6 and Ana2, and the Ana2 CCCD tetramer structure provides important information on how these proteins might cooperate to form a cartwheel structure.

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Author details

  1. Matthew A Cottee

    Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.

  2. Nadine Muschalik

    Division of Cell Biology, MRC-Laboratory of Molecular Biology, Cambridge, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.

  3. Steven Johnson

    Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.

  4. Joanna Leveson

    Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.

  5. Jordan W Raff

    Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom
    For correspondence
    jordan.raff@path.ox.ac.uk
    Competing interests
    The authors declare that no competing interests exist.

  6. Susan M Lea

    Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.

Copyright

© 2015, Cottee et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Matthew A Cottee
  2. Nadine Muschalik
  3. Steven Johnson
  4. Joanna Leveson
  5. Jordan W Raff
  6. Susan M Lea
(2015)
The homo-oligomerisation of both Sas-6 and Ana2 is required for efficient centriole assembly in flies
eLife 4:e07236.
https://doi.org/10.7554/eLife.07236

Share this article

https://doi.org/10.7554/eLife.07236

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    1. Structural Biology and Molecular Biophysics
    2. Cell Biology

    The Caenorhabditis elegans protein SAS-5 forms large oligomeric assemblies critical for centriole formation

    Kacper B Rogala, Nicola J Dynes ... Ioannis Vakonakis
    Research Article Updated

    Centrioles are microtubule-based organelles crucial for cell division, sensing and motility. In Caenorhabditis elegans, the onset of centriole formation requires notably the proteins SAS-5 and SAS-6, which have functional equivalents across eukaryotic evolution. Whereas the molecular architecture of SAS-6 and its role in initiating centriole formation are well understood, the mechanisms by which SAS-5 and its relatives function is unclear. Here, we combine biophysical and structural analysis to uncover the architecture of SAS-5 and examine its functional implications in vivo. Our work reveals that two distinct self-associating domains are necessary to form higher-order oligomers of SAS-5: a trimeric coiled coil and a novel globular dimeric Implico domain. Disruption of either domain leads to centriole duplication failure in worm embryos, indicating that large SAS-5 assemblies are necessary for function in vivo.