Temporal and spatial regulation of phosphoinositide signaling mediates cytokinesis.

Polarity is a prominent feature of both chemotaxis and cytokinesis. In chemotaxis, polarity is established by local accumulation of PI(3,4,5)P3 at the cell's leading edge, achieved through temporal and spatial regulation of PI3 kinases and the tumor suppressor, PTEN. We find that as migrating D. discoideum cells round up to enter cytokinesis, PI(3,4,5)P3 signaling is uniformly suppressed. Then, as the spindle and cell elongate, PI3 kinases and PTEN move to and function at the poles and furrow, respectively. Cell lines lacking both of these enzymatic activities fail to modulate PI(3,4,5)P3 levels, are defective in cytokinesis, and cannot divide in suspension. The cells continue to grow and duplicate their nuclei, generating large multinucleate cells. Furrows that fail to ingress between nuclei are unable to stably accumulate myosin filaments or suppress actin-filled ruffles. We propose that phosphoinositide-linked circuits, similar to those that bring about asymmetry during cell migration, also regulate polarity in cytokinesis.

[1]  W. Sellers,et al.  The PTEN tumor suppressor protein: an antagonist of phosphoinositide 3-kinase signaling. , 2000, Biochimica et biophysica acta.

[2]  T. Meyer,et al.  Spatial Sensing in Fibroblasts Mediated by 3′ Phosphoinositides , 2000, The Journal of cell biology.

[3]  J. Ptak,et al.  High Frequency of Mutations of the PIK3CA Gene in Human Cancers , 2004, Science.

[4]  F. Haluska,et al.  PTEN signaling pathways in melanoma , 2003, Oncogene.

[5]  D. Drubin,et al.  Functional cooperation between the microtubule and actin cytoskeletons. , 2000, Current opinion in cell biology.

[6]  C. Parent,et al.  A cell's sense of direction. , 1999, Science.

[7]  Paul Herzmark,et al.  Lipid products of PI(3)Ks maintain persistent cell polarity and directed motility in neutrophils , 2002, Nature Cell Biology.

[8]  J. Schwartz,et al.  Microtubule-mediated centrosome motility and the positioning of cleavage furrows in multinucleate myosin II-null cells. , 1998, Journal of cell science.

[9]  R. Firtel,et al.  A novel Akt/PKB-related kinase is essential for morphogenesis in Dictyostelium , 2000, Current Biology.

[10]  D. Guertin,et al.  Cytokinesis in Eukaryotes , 2002, Microbiology and Molecular Biology Reviews.

[11]  P. Devreotes,et al.  Eukaryotic Chemotaxis: Distinctions between Directional Sensing and Polarization* , 2003, Journal of Biological Chemistry.

[12]  G. Gerisch,et al.  Chemoattractant-controlled accumulation of coronin at the leading edge of Dictyostelium cells monitored using a green fluorescent protein–coronin fusion protein , 1995, Current Biology.

[13]  I. Weber,et al.  Cytoskeletal protein mutations and cell motility in Dictyostelium. , 1999, Biochemical Society symposium.

[14]  T. Uyeda,et al.  Myosin II-independent cytokinesis in Dictyostelium: its mechanism and implications. , 2000, Cell structure and function.

[15]  E. Martz.  Finishing the job. , 2006, Delaware medical journal.

[16]  D A Knecht,et al.  Antisense RNA inactivation of myosin heavy chain gene expression in Dictyostelium discoideum. , 1987, Science.

[17]  Francisca Vazquez,et al.  Phosphorylation of the PTEN Tail Regulates Protein Stability and Function , 2000, Molecular and Cellular Biology.

[18]  P. Iglesias,et al.  Chemoattractant-induced phosphatidylinositol 3,4,5-trisphosphate accumulation is spatially amplified and adapts, independent of the actin cytoskeleton , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[19]  J. Spudich,et al.  Towards a molecular understanding of cytokinesis. , 2000, Trends in cell biology.

[20]  S. Labeit,et al.  Towards a molecular understanding of titin. , 1992, The EMBO journal.

[21]  K. Oegema,et al.  Rappaport rules: cleavage furrow induction in animal cells. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[22]  P. Iglesias,et al.  Two complementary, local excitation, global inhibition mechanisms acting in parallel can explain the chemoattractant-induced regulation of PI(3,4,5)P3 response in dictyostelium cells. , 2004, Biophysical journal.

[23]  N. Fuse,et al.  Heterotrimeric G Proteins Regulate Daughter Cell Size Asymmetry in Drosophila Neuroblast Divisions , 2003, Current Biology.

[24]  R. Gräf,et al.  Regulated expression of the centrosomal protein DdCP224 affects microtubule dynamics and reveals mechanisms for the control of supernumerary centrosome number. , 2003, Molecular biology of the cell.

[25]  G. Gerisch,et al.  Patterns of cellular activities based on protein sorting in cell motility, endocytosis and cytokinesis. , 1999, Biochemical Society symposium.

[26]  M. Balasubramanian,et al.  Cytokinesis: relative alignment of the cell division apparatus and the mitotic spindle. , 2003, Current opinion in cell biology.

[27]  Jonathon Howard,et al.  The Distribution of Active Force Generators Controls Mitotic Spindle Position , 2003, Science.

[28]  N. Masuyama,et al.  Akt mediates Rac/Cdc42-regulated cell motility in growth factor-stimulated cells and in invasive PTEN knockout cells , 2001, Current Biology.

[29]  P. Devreotes,et al.  Two phases of actin polymerization display different dependencies on PI(3,4,5)P3 accumulation and have unique roles during chemotaxis. , 2003, Molecular biology of the cell.

[30]  Mimi Shirasu-Hiza,et al.  Dynamics of the mitotic spindle--potential therapeutic targets. , 2003, Progress in cell cycle research.

[31]  Yu-Li Wang,et al.  Distinct roles of the equatorial and polar cortices in the cleavage of adherent cells , 2001, Current Biology.

[32]  Y. Fukui,et al.  Reorganization of microtubules during mitosis in Dictyostelium: Dissociation from MTOC and selective assembly/disassembly in situ , 1987 .

[33]  Francisca Vazquez,et al.  Novel Mechanism of PTEN Regulation by Its Phosphatidylinositol 4,5-Bisphosphate Binding Motif Is Critical for Chemotaxis* , 2004, Journal of Biological Chemistry.

[34]  R. Firtel,et al.  Receptor-mediated regulation of PI3Ks confines PI(3,4,5)P3 to the leading edge of chemotaxing cells. , 2003, Molecular biology of the cell.

[35]  C. Mitchell,et al.  Increased levels of phosphatidylinositol 3‐kinase activity in colorectal tumors , 1998, Cancer.

[36]  A. Hyman,et al.  A requirement for Rho and Cdc42 during cytokinesis in Xenopus embryos , 1997, Current Biology.

[37]  P. Devreotes,et al.  Tumor Suppressor PTEN Mediates Sensing of Chemoattractant Gradients , 2002, Cell.

[38]  M. Mansukhani,et al.  Reduced expression of PTEN correlates with breast cancer progression. , 2002, Human pathology.

[39]  C. Rieder,et al.  Mitosis in vertebrate somatic cells with two spindles: implications for the metaphase/anaphase transition checkpoint and cleavage. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[40]  C. Parent,et al.  Localization of the G Protein βγ Complex in Living Cells During Chemotaxis , 2000 .

[41]  P. Devreotes,et al.  Chemotaxis in eukaryotic cells: a focus on leukocytes and Dictyostelium. , 1988, Annual review of cell biology.

[42]  Y. Fukui.  Real‐time high‐resolution optical sectioning suggests biphasic cytokinetic mechanism in Dictyostelium discoideum , 2000, Microscopy research and technique.

[43]  M. Chung,et al.  Inactivation of the PTEN gene protein product is associated with the invasiveness and metastasis, but not angiogenesis, of breast cancer , 2004, Pathology international.

[44]  R. Gräf,et al.  Centrosomal microtubule plus end tracking proteins and their role in Dictyostelium cell dynamics , 2004, Journal of Muscle Research & Cell Motility.

[45]  Masayuki Yamamoto,et al.  Type II Myosin Heavy Chain Encoded by the myo2 Gene Composes the Contractile Ring during Cytokinesis in Schizosaccharomyces pombe , 1997, The Journal of cell biology.

[46]  Richard A. Firtel,et al.  Spatial and Temporal Regulation of 3-Phosphoinositides by PI 3-Kinase and PTEN Mediates Chemotaxis , 2002, Cell.

[47]  L. Cantley,et al.  A novel phosphatidylinositol(3,4,5)P3 pathway in fission yeast , 2004, The Journal of cell biology.

[48]  T. Sasaki,et al.  Regulation of cytoplasmic division of Xenopus embryo by rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI) , 1993, The Journal of cell biology.

[49]  R. Rappaport,et al.  Establishment of the mechanism of cytokinesis in animal cells. , 1986, International review of cytology.

[50]  P. Devreotes,et al.  Temporal and spatial regulation of chemotaxis. , 2002, Developmental cell.

[51]  C. D’Souza-Schorey,et al.  Finishing the job: cytoskeletal and membrane events bring cytokinesis to an end. , 2004, Experimental cell research.

[52]  S. Wheatley,et al.  Midzone microtubule bundles are continuously required for cytokinesis in cultured epithelial cells [published erratum appears in J Cell Biol 1996 Dec;135(6 Pt 1):1679] , 1996, The Journal of cell biology.

[53]  C. Sawyers,et al.  The phosphatidylinositol 3-Kinase–AKT pathway in human cancer , 2002, Nature Reviews Cancer.

[54]  J. Spudich,et al.  Disruption of the Dictyostelium myosin heavy chain gene by homologous recombination. , 1987, Science.

[55]  B. Baum,et al.  Myosin II-Dependent Cortical Movement Is Required for Centrosome Separation and Positioning during Mitotic Spindle Assembly , 2004, Cell.

[56]  K. Zhou,et al.  A phosphatidylinositol (PI) kinase gene family in Dictyostelium discoideum: biological roles of putative mammalian p110 and yeast Vps34p PI 3-kinase homologs during growth and development , 1995, Molecular and cellular biology.

[57]  Daniel J. Lew,et al.  Involvement of an Actomyosin Contractile Ring in Saccharomyces cerevisiae Cytokinesis , 1998, The Journal of cell biology.

[58]  C. Parent,et al.  Localization of the G protein betagamma complex in living cells during chemotaxis. , 2000, Science.

[59]  J. Spudich,et al.  A mitotic kinesin-like protein required for normal karyokinesis, myosin localization to the furrow, and cytokinesis in Dictyostelium. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[60]  J. Canman,et al.  Microtubules suppress actomyosin-based cortical flow in Xenopus oocytes. , 1997, Journal of cell science.

[61]  Rong Li,et al.  Sequential Assembly of Myosin II, an IQGAP-like Protein, and Filamentous Actin to a Ring Structure Involved in Budding Yeast Cytokinesis , 1998, The Journal of cell biology.

[62]  Douglas N Robinson,et al.  Mechanics and regulation of cytokinesis. , 2004, Current opinion in cell biology.

[63]  C. Huang,et al.  Roles of PI3K in neutrophil function. , 2004, Current topics in microbiology and immunology.

[64]  Y. Wang,et al.  Signals from the spindle midzone are required for the stimulation of cytokinesis in cultured epithelial cells. , 1996, Molecular biology of the cell.

[65]  Richard A. Firtel,et al.  Role of Phosphatidylinositol 3′ Kinase and a Downstream Pleckstrin Homology Domain–Containing Protein in Controlling Chemotaxis inDictyostelium , 2001, The Journal of cell biology.

[66]  Hui Ma,et al.  Chemoattractant‐mediated transient activation and membrane localization of Akt/PKB is required for efficient chemotaxis to cAMP in Dictyostelium , 1999, The EMBO journal.

[67]  J. A. Brill,et al.  A phospholipid kinase regulates actin organization and intercellular bridge formation during germline cytokinesis. , 2000, Development.

[68]  M. Schleicher,et al.  Dictyostelium discoideum: a genetic model system for the study of professional phagocytes. Profilin, phosphoinositides and the lmp gene family in Dictyostelium. , 2001, Biochimica et biophysica acta.

[69]  D. Manning.  Evidence Mounts for Receptor-Independent Activation of Heterotrimeric G Proteins Normally in Vivo: Positioning of the Mitotic Spindle in C. Elegans , 2003, Science's STKE.

[70]  D. Goberdhan,et al.  PTEN: tumour suppressor, multifunctional growth regulator and more. , 2003, Human molecular genetics.

[71]  R. Mamillapalli,et al.  Genetic deletion of the Pten tumor suppressor gene promotes cell motility by activation of Rac1 and Cdc42 GTPases , 2000, Current Biology.