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

Centre for Cancer Biomedicine, Faculty of Medicine, University of Oslo, Montebello, N-0310 Oslo, Norway.
Nature Cell Biology (Impact Factor: 19.68). 03/2010; 12(4):362-71. DOI: 10.1038/ncb2036
Source: PubMed


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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    • "In addition, PI(3)P is also central to autophagy in mammalian cells (Petiot et al., 2000) and yeast (Obara and Ohsumi, 2011). More recently, its role in cytokinesis has been demonstrated (Sagona et al., 2010). PI(3)P exercises its effects by recruiting a variety of cytosolic proteins that contain mainly FYVE (FAB1-YOTB-Vac1-EEA1) domains (Kutateladze et al., 1999; Katzmann et al., 2003; Hayakawa et al., 2004) or PX domains (Cheever et al., 2001). "
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    ABSTRACT: Phosphoinositides regulate numerous cellular processes, by recruiting cytosolic effector proteins and acting as membrane signaling entities. The cellular metabolism and localization of phosphoinositides are tightly regulated by distinct lipid kinases and phosphatases. Here, we identify and characterize a unique phosphatidylinositol 3-Kinase (PI3K) in Toxoplasma gondii, a protozoan parasite belonging to the phylum Apicomplexa. Conditional depletion of this enzyme and subsequently of its product, PI(3)P, drastically alters the morphology and inheritance of the apicoplast, an endosymbiontic organelle of algal origin that is a unique feature of many Apicomplexa. We searched the T. gondii genome for PI(3)P binding proteins and identified in total six PX and FYVE-domain containing proteins including a PIKfyve lipid kinase, which phosphorylates PI(3)P into PI(3,5)P2 . While depletion of putative PI(3)P binding proteins shows that they are not essential for parasite growth and apicoplast biology, conditional disruption of PIKfyve induces enlarged apicoplasts, as observed upon the loss of PI(3)P. A similar defect of apicoplast homeostasis was also observed by knocking-down the PIKfyve regulatory protein ArPIKfyve, suggesting that in T. gondii, PI(3)P-related function for the apicoplast might mainly be to serve as a precursor for the synthesis of PI(3,5)P2 . Accordingly, PI3K is conserved in all apicomplexan parasites whereas PIKfyve and ArPIKfyve are absent in Cryptosporidium species which lack an apicoplast, supporting a direct role of PI(3,5)P2 in apicoplast homeostasis. This study enriches the already diverse functions attributed to PI(3,5)P2 in eukaryotic cells and highlights these parasite lipid kinases as potential drug targets.
    Cellular Microbiology 10/2014; 17(4). DOI:10.1111/cmi.12383 · 4.92 Impact Factor
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    • "First, the protein and lipid composition of the plasma membrane is remodeled, which involves vesicular delivery through several parallel trafficking pathways [e.g. Rab35, Rab11-FIP3 (also known as Rab11 family interacting protein 3), phosphatidylinositol 3-phosphate (PI3P), exocyst] (Echard, 2012b; Fielding et al., 2005; Goss and Toomre, 2008; Gromley et al., 2005; Kouranti et al., 2006; Montagnac et al., 2008; Neto et al., 2011; Sagona et al., 2010; Wilson et al., 2005). For instance, the Rab11-and the Rab35-regulated pathways prevent F-actin accumulation at the plasma membrane, and are both required for normal abscission (Chesneau et al., 2012; Dambournet et al., 2011; Echard, 2012a; Schiel et al., 2012). "
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    ABSTRACT: The midbody remnant (MBR) that is generated after cytokinesis abscission has recently attracted a lot of attention, since it may have crucial consequences for cell differentiation and tumorigenesis in mammalian cells. In those cells, it has been reported that the MBR is either released into the extracellular medium, or retracted into one of the two daughter cells where it can be degraded by autophagy. Here, we describe a major alternative pathway in a variety of human and mouse immortalized/cancer and primary stem cells. Using correlative light/scanningEM microscopy and quantitative assays, we found that sequential abscissions on both sides of the midbody generate free MBRs, which are tightly associated to the cell surface through a Ca(++)/Mg(++)-dependent receptor. Surprisingly, MBRs move over the cell surface for several hours, before being eventually engulfed by an actin-dependent phagocytosis-like mechanism. Mathematical modelling combined to experiments further demonstrates that lysosomal activities fully account for clearance of MBRs after engulfment. This study changes our vision of how MBRs are inherited and degraded in mammalian cells, and suggests a mechanism by which MBRs might signal over long distances between cells.
    Journal of Cell Science 07/2014; 127(17). DOI:10.1242/jcs.154732 · 5.43 Impact Factor
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    • "PI3K-III has been shown to form complexes that regulate cytokinesis, and that are very similar to the autophagy-related PI3K-III complexes. Downregulation of various components of the PI3K-III complex resulted in impaired cytokinesis [50,51]. It is therefore possible that cytokinesis in our mutant is inhibited either as a result of impaired autophagy, whereby the VMP1 deficiency is solely the cause for said impairment; or that the VMP1 deficiency causes the cytokinesis defect in a more direct fashion, possibly through the disruption of PI3K-III complexes. "
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    ABSTRACT: Background The versatile Vacuole Membrane Protein 1 (VMP1) has been previously investigated in six species. It has been shown to be essential in macroautophagy, where it takes part in autophagy initiation. In addition, VMP1 has been implicated in organellar biogenesis; endo-, exo- and phagocytosis, and protein secretion; apoptosis; and cell adhesion. These roles underly its proven involvement in pancreatitis, diabetes and cancer in humans. Results In this study we analyzed a VMP1 homologue from the green alga Chlamydomonas reinhardtii. CrVMP1 knockdown lines showed severe phenotypes, mainly affecting cell division as well as the morphology of cells and organelles. We also provide several pieces of evidence for its involvement in macroautophagy. Conclusion Our study adds a novel role to VMP1's repertoire, namely the regulation of cytokinesis. Though the directness of the observed effects and the mechanisms underlying them remain to be defined, the protein's involvement in macroautophagy in Chlamydomonas, as found by us, suggests that CrVMP1 shares molecular characteristics with its animal and protist counterparts.
    BMC Plant Biology 05/2014; 14(1):121. DOI:10.1186/1471-2229-14-121 · 3.81 Impact Factor
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