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Myosin 1b promotes the formation of post-Golgi carriers by regulating actin assembly and membrane remodelling at the trans-Golgi network

Nature Cell Biology volume 13, pages 779–789 (2011)Cite this article

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Abstract

The function of organelles is intimately associated with rapid changes in membrane shape. By exerting force on membranes, the cytoskeleton and its associated motors have an important role in membrane remodelling. Actin and myosin 1 have been implicated in the invagination of the plasma membrane during endocytosis. However, whether myosin 1 and actin contribute to the membrane deformation that gives rise to the formation of post-Golgi carriers is unknown. Here we report that myosin 1b regulates the actin-dependent post-Golgi traffic of cargo, generates force that controls the assembly of F-actin foci and, together with the actin cytoskeleton, promotes the formation of tubules at the TGN. Our results provide evidence that actin and myosin 1 regulate organelle shape and uncover an important function for myosin 1b in the initiation of post-Golgi carrier formation by regulating actin assembly and remodelling TGN membranes.

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Figure 1: Distribution of Myo1b, MPR, F-actin and the Arp2/3 complex in the perinuclear region.
Figure 2: Myo1b controls the steady-state distribution of MPR.
Figure 3: Myo1b knockdown impairs the TGN exit of cargo.
Figure 4: Myo1b controls the formation of tubular carriers at the TGN.
Figure 5: Myo1b depletion reduces the number of F-actin foci and of Arp2/3 complex structures in the vicinity of the TGN.
Figure 6: Functional Myo1b is required for organizing F-actin–Arp2/3 foci.
Figure 7: Depletion of the Arp2/3 complex induces MPR accumulation in TGN and inhibits post-Golgi carrier formation.
Figure 8: Model for the role of Myo1b in the formation of tubular carriers at the TGN.

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Acknowledgements

We thank L. Salas-Cortes for designing the Myo1b siRNA, M. Prospéri for Myo1b antibodies and the Cherry–Myo1b, FlagHA–Myo1b-5MR and FlagHA–Myo1b-5ME constructs and J. Lee-Tin-Wah and P. Martin for helping set up the actin sliding assay. We thank V. Fraisier, J-B. Sibarita, F. Waharte and L. Sengmanivong for their expertise in microscopy and the Nikon imaging centre@Institut Curie-CNRS. We thank E. Derivery for setting up the Myo1b purification with the FlipIn system. We thank J. Kean for critical reading of the manuscript.

This work has been supported by the Institut Curie, the CNRS and the Agence Nationale pour la Recherche (grant ANR 09-BLAN-0027). C.G.A. has been the recipient of an EMBO long-term fellowship (ALTF 607-2006) and a Marie Curie action intra-European fellowship for career development (FP7-PEOPLE-2007-2-1-IEF N. 2200088).

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Authors and Affiliations

  1. Institut Curie, Centre de Recherche, Paris, F-75248, France

    Claudia G. Almeida, Ayako Yamada, Danièle Tenza, Daniel Louvard, Graça Raposo & Evelyne Coudrier

  2. Morphogenesis and Cell Signalization CNRS, UMR144, Paris, F-75248, France

    Claudia G. Almeida, Daniel Louvard & Evelyne Coudrier

  3. Cell and Tissue Imaging Facility, CNRS UMR 144, Paris F-75248, France

    Ayako Yamada

  4. Membrane and Cell Functions, CNRS UMR 168, Paris F-75248, France

    Danièle Tenza & Graça Raposo

  5. Structure and Membrane Compartments CNRS, UMR144, Paris, F-75248, France

    Danièle Tenza & Graça Raposo

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Contributions

C.G.A. and E.C. conceived the project and wrote the manuscript; C.G.A. generated and analysed most of the data; A.Y. carried out Myo1b and Myo1b mutant purification as well as their characterization in vitro; D.T. and G.R. generated and analysed the electron microscopy data; D.L. revised the manuscript.

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Correspondence to Evelyne Coudrier.

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Almeida, C., Yamada, A., Tenza, D. et al. Myosin 1b promotes the formation of post-Golgi carriers by regulating actin assembly and membrane remodelling at the trans-Golgi network. Nat Cell Biol 13, 779–789 (2011). https://doi.org/10.1038/ncb2262

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