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Rab35 GTPase and OCRL phosphatase remodel lipids and F-actin for successful cytokinesis

Nature Cell Biology volume 13, pages 981–988 (2011)Cite this article

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Abstract

Abscission is the least understood step of cytokinesis. It consists of the final cut of the intercellular bridge connecting the sister cells at the end of mitosis, and is thought to involve membrane trafficking as well as lipid and cytoskeleton remodelling1,2,3,4,5,6. We previously identified the Rab35 GTPase as a regulator of a fast recycling endocytic pathway that is essential for post-furrowing cytokinesis stages7. Here, we report that the phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2) 5-phosphatase OCRL, which is mutated in Lowe syndrome patients8,9, is an effector of the Rab35 GTPase in cytokinesis abscission. GTP-bound (active) Rab35 directly interacts with OCRL and controls its localization at the intercellular bridge. Depletion of Rab35 or OCRL inhibits cytokinesis abscission and is associated with local abnormal PtdIns(4,5)P2 and F-actin accumulation in the intercellular bridge. These division defects are also found in cell lines derived from Lowe patients and can be corrected by the addition of low doses of F-actin depolymerization drugs. Our data demonstrate that PtdIns(4,5)P2 hydrolysis is important for normal cytokinesis abscission to locally remodel the F-actin cytoskeleton in the intercellular bridge. They also reveal an unexpected role for the phosphatase OCRL in cell division and shed new light on the pleiotropic phenotypes associated with Lowe disease.

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Figure 1: OCRL directly interacts with GTP-bound Rab35.
Figure 2: Rab35 activation controls the accumulation of an OCRL pool in the cytokinesis intercellular bridge.
Figure 3: Cytokinesis abscission defects in cells depleted for Rab35 or OCRL, and in Lowe syndrome patient cells.
Figure 4: Abnormal local accumulation of PtdIns(4,5)P2 and F-actin at the intercellular bridges in cells depleted for OCRL or Rab35 and in Lowe syndrome patient cells.
Figure 5: Abscission defects observed in cells depleted for OCRL or Rab35 and in Lowe syndrome patient cells are suppressed by low doses of F-actin depolymerizing drugs.

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Acknowledgements

We thank R. Woscholski (Imperial College, London, UK), O. Dorseuil (Institut Cochin, Paris, France), A. Gautreau (CNRS Lebs, Gif, France), A. Alcover (Institut Pasteur, Paris), S. Miserey-Lenkei, P. Benaroch and F. Nagano (Institut Curie, Paris, France) for providing reagents and plasmids; V. Chauvet, A. Gautreau, A. Houdusse, B. Payrastre, N. Vitale and R. Weil for helpful discussions; S. Miserey-Lenkei and E. Crowell for critical reading of the manuscript; F. Legendre (Hôpital Necker, Paris, France) for the establishment of patients cell lines; and J. Lunardi (CHU Grenoble, France) and G. Baujat for the pediatric network. We thank the ‘Association du Syndrome de Lowe’, patients and parents. We thank the Plate-Forme d’Imagerie Dynamique (PFID) and Imagopole, Institut Pasteur, for microscopes and assistance. The authors thank V. Fraisier, L. Sengmanivong, J-B. Sibarita and J. Salamero for support in microscopy, and acknowledge the Nikon Imaging Centre at Institut Curie-CNRS. This work has been supported by the Institut PASTEUR (G5 program), the Institut CURIE, the CNRS, the INSERM, the Agence Nationale pour la Recherche (grants ANR-Maladies Rares, GIS-Maladies Rares to B.G. and R.S., and ANR 07-JCJC-0089 to A.E.) and the Schlumberger Foundation for Education and Research—FSER (A.E.). D.D. and M.M. have been supported by the Ministère de la Recherche et de l’Enseignement Supérieur, and D.D. and L.C. have been supported by the Association pour la Recherche sur le Cancer.

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Authors and Affiliations

  1. Membrane Traffic and Cell Division Lab. 25–28 rue du Dr Roux, Institut Pasteur, 75724 Paris cedex 15, France

    Daphné Dambournet, Mickael Machicoane, Laurent Chesneau, Murielle Rocancourt & Arnaud Echard

  2. CNRS URA2582, France

    Daphné Dambournet, Mickael Machicoane, Laurent Chesneau, Murielle Rocancourt & Arnaud Echard

  3. Institut Pasteur, Imagopole, Plate-forme de microscopie ultrastructurale, 25–28 rue du Dr Roux, 75724 Paris cedex 15, France

    Martin Sachse

  4. Institut Curie, Molecular Mechanisms of Intracellular Transport Lab. 25 rue d’Ulm, 75005 Paris, France

    Ahmed El Marjou & Bruno Goud

  5. CNRS UMR144, France

    Ahmed El Marjou & Bruno Goud

  6. Hybrigenics SA, 3–5 impasse Reille, 75014 Paris, France

    Etienne Formstecher

  7. AP-HP Hôpital Necker, Service de Néphrologie Pédiatrique, Inserm U983, Paris, France

    Rémi Salomon

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Contributions

D.D., M.M., L.C., M.S. and A.E. designed and analysed the experiments; D.D., M.M., L.C., M.S., M.R., A.E.M., E.F. and A.E. did the experimental work; R.S. and B.G. provided reagents; D.D., M.M., L.C., M.S., B.G. and A.E. wrote or edited the manuscript.

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Correspondence to Arnaud Echard.

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Dambournet, D., Machicoane, M., Chesneau, L. et al. Rab35 GTPase and OCRL phosphatase remodel lipids and F-actin for successful cytokinesis. Nat Cell Biol 13, 981–988 (2011). https://doi.org/10.1038/ncb2279

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