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PtdIns(3)P controls cytokinesis through KIF13A-mediated recruitment of FYVE-CENT to the midbody

Nature Cell Biology volume 12, pages 362–371 (2010)Cite this article

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Abstract

Several subunits of the class III phosphatidylinositol-3-OH kinase (PI(3)K-III) complex are known as tumour suppressors. Here we uncover a function for this complex and its catalytic product phosphatidylinositol-3-phosphate (PtdIns(3)P) in cytokinesis. We show that PtdIns(3)P localizes to the midbody during cytokinesis and recruits a centrosomal protein, FYVE-CENT (ZFYVE26), and its binding partner TTC19, which in turn interacts with CHMP4B, an endosomal sorting complex required for transport (ESCRT)-III subunit implicated in the abscission step of cytokinesis. Translocation of FYVE-CENT and TTC19 from the centrosome to the midbody requires another FYVE-CENT-interacting protein, the microtubule motor KIF13A. Depletion of the VPS34 or Beclin 1 subunits of PI(3)K-III causes cytokinesis arrest and an increased number of binucleate and multinucleate cells, in a similar manner to the depletion of FYVE-CENT, KIF13A or TTC19. These results provide a mechanism for the translocation and docking of a cytokinesis regulatory machinery at the midbody.

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Figure 1: PtdIns(3)P localizes to the midbody and is required for cytokinesis.
Figure 2: siRNA screening for PtdIns(3)P-binding proteins regulating early cytokinesis.
Figure 3: FYVE-CENT localizes to the centrosome and midbody.
Figure 4: FYVE-CENT is required for cytokinesis.
Figure 5: A functional FYVE domain is important for the targeting of FYVE-CENT to the midbody.
Figure 6: FYVE-CENT binds directly to TTC19 and KIF13A.
Figure 7: The FYVE-CENT-interacting proteins KIF13A and TTC19 localize to the centrosome and midbody and control cytokinesis.
Figure 8: The interplay between FYVE-CENT, KIF13A and TTC19 controls cytokinesis.

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Acknowledgements

We thank Nobutaka Hirokawa for providing the expression construct for Myc–KIF13A, Aviva Tolkovsky for the gift of HeLa GFP–LC3 cells, Wes Sundquist for providing CHMP4A constructs, and Thomas P. Neufeld for the vps34 mutant flies. A.P.S. is a PhD student of the ENDOCYTE Research Training Network of the European Union. I.P.N. is a postdoctoral fellow of the Stem Cell Research Programme of the Research Council of Norway. N.M.P. is a postdoctoral fellow of the South-Eastern Norway Regional Health Authority. C.R. is a postdoctoral fellow, and T.E.R. and R.I.S. are senior research fellows, of the Norwegian Cancer Society.

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Authors and Affiliations

  1. Centre for Cancer Biomedicine, Faculty of Medicine, University of Oslo, Montebello, Oslo, N-0310, Norway

    Antonia P. Sagona, Ioannis P. Nezis, Nina Marie Pedersen, Knut Liestøl, John Poulton, Tor Erik Rusten, Rolf I. Skotheim, Camilla Raiborg & Harald Stenmark

  2. Department of Biochemistry Institute for Cancer Research, The Norwegian Radium Hospital, Oslo University Hospital, Montebello, Oslo, N-0310, Norway

    Antonia P. Sagona, Ioannis P. Nezis, Nina Marie Pedersen, John Poulton, Tor Erik Rusten, Camilla Raiborg & Harald Stenmark

  3. Department of Cancer Prevention, Institute for Cancer Research, The Norwegian Radium Hospital, Oslo University Hospital, Montebello, Oslo, N-0310, Norway

    Rolf I. Skotheim

  4. Department of Informatics, University of Oslo, Montebello, Oslo, N-0310, Norway

    Knut Liestøl

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  1. Antonia P. Sagona
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Contributions

A.P.S. designed and performed cloning, transfection, confocal fluorescence microscopy, siRNA knock-down, rescue, immunoblotting, pull-down and liposome binding assay experiments and contributed to the writing of the manuscript. I.P.N. conceived and designed experiments, performed electron microscopy experiments, observed and analysed data from confocal fluorescence microscopy experiments and wrote the draft manuscript. N.M.P. performed transfection, co-immunoprecipitation and pull-down experiments, participated in siRNA screening and reviewed the manuscript. K.L. performed statistical analyses and edited the manuscript. J.P. and T.E.R. conceived and performed studies of Drosophila vps34 mutants and edited the manuscript. R.I.S. reviewed cancer data and edited the manuscript. C.R. performed siRNA screening and cytokinesis analyses and edited the manuscript. H.S. conceived and designed experiments, supervised the study and wrote the final version of the manuscript.

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Correspondence to Harald Stenmark.

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Sagona, A., Nezis, I., Pedersen, N. et al. PtdIns(3)P controls cytokinesis through KIF13A-mediated recruitment of FYVE-CENT to the midbody. Nat Cell Biol 12, 362–371 (2010). https://doi.org/10.1038/ncb2036

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