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Anthrax toxin receptor 2a controls mitotic spindle positioning

Nature Cell Biology volume 15, pages 28–39 (2013)Cite this article

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Abstract

Oriented mitosis is essential during tissue morphogenesis. The Wnt/planar cell polarity (Wnt/PCP) pathway orients mitosis in a number of developmental systems, including dorsal epiblast cell divisions along the animal–vegetal (A–V) axis during zebrafish gastrulation. How Wnt signalling orients the mitotic plane is, however, unknown. Here we show that, in dorsal epiblast cells, anthrax toxin receptor 2a (Antxr2a) accumulates in a polarized cortical cap, which is aligned with the embryonic A–V axis and forecasts the division plane. Filamentous actin (F-actin) also forms an A–V polarized cap, which depends on Wnt/PCP and its effectors RhoA and Rock2. Antxr2a is recruited to the cap by interacting with actin. Antxr2a also interacts with RhoA and together they activate the diaphanous-related formin zDia2. Mechanistically, Antxr2a functions as a Wnt-dependent polarized determinant, which, through the action of RhoA and zDia2, exerts torque on the spindle to align it with the A–V axis.

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Figure 1: Antxr2a and F-actin are polarized at the cell cortex and form caps in dividing dorsal epiblast cells.
Figure 2: Antxr2a/F-actin caps align with the embryonic A–V axis and forecast the plane of mitosis.
Figure 3: The mitotic spindle aligns to the embryonic A–V axis through episodes of directed rotation.
Figure 4: RhoA interacts with Antxr2a and is required for Antxr2a/F-actin cap alignment.
Figure 5: Spindle rotation phenotype of zdia2 and antxr2a morphants.
Figure 6: The actin–myosin cytoskeleton is required for proper alignment of the Antxr2a/F-actin caps.
Figure 7: PCP pathway-dependent alignment of the F-actin and Antxr2a caps along the embryonic axis.
Figure 8: Phenotypic summary and a model for spindle alignment and oriented mitosis in dorsal epiblast cells.

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Acknowledgements

We are grateful to B. Kunz and N. Zangger for generating the zAntxr2a construct for the pulldown, the phylogenetic tree and multiple sequence alignments, S. Abke, E. Paluch, C. Low, G. Weidinger, R. W. Köster, E. Kardash, L. Solnica-Krezel, A. J. Ridley, G. Davidson, S. Loubéry and O. Schaad, and the bioimaging and fish platforms for reagents and technical help and T. Hannich for critical reading. This work was supported by the SNSF, the Swiss SystemsX.ch initiative and LipidX-2008/011 (M.G-G. and F.G.v.d.G.), by the Fondation SANTE-Vaduz/Aide au Soutien des Nouvelles Thérapies (F.G.v.d.G.) and by the ERC, the NCCR Frontiers in Genetics and Chemical Biology programmes and the Polish–Swiss research program (M.G-G.).

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Authors and Affiliations

  1. Departments of Biochemistry and Molecular Biology, Sciences II, 30 Quai Ernest-Ansermet, CH-1211 Geneva 4, Switzerland

    I. Castanon, L. Holtzer & M. González-Gaitán

  2. Global Health Institute, Ecole Polytechnique Fédérale de Lausanne, Station 15, CH-1015 Lausanne, Switzerland

    L. Abrami & F. G. van der Goot

  3. Institute of Science and Technology Austria, Am Campus 1, A-3400 Klosterneuburg, Austria

    C. P. Heisenberg

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Contributions

I.C. designed and carried out the experiments, generated and interpreted most of the data and prepared the manuscript. L.A. designed and carried out the immunoprecipitation experiments. L.H. designed the algorithm and simulation assays for the directed rotation experiments and C.P.H. supported experimentation and contributed to data interpretation. I.C., F.G.v.d.G. and M.G-G. wrote the manuscript.

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Correspondence to F. G. van der Goot or M. González-Gaitán.

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Castanon, I., Abrami, L., Holtzer, L. et al. Anthrax toxin receptor 2a controls mitotic spindle positioning. Nat Cell Biol 15, 28–39 (2013). https://doi.org/10.1038/ncb2632

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