Regulation of mitotic spindle orientation: an integrated view

Mitotic spindle orientation is essential for cell fate decisions, epithelial maintenance, and tissue morphogenesis. In most animal cell types, the dynein motor complex is anchored at the cell cortex and exerts pulling forces on astral microtubules to position the spindle. Early studies identified the evolutionarily conserved Gαi/LGN/NuMA complex as a key regulator that polarizes cortical force generators. In recent years, a combination of genetics, biochemistry, modeling, and live imaging has contributed to decipher the mechanisms of spindle orientation. Here, we highlight the dynamic nature of the assembly of this complex and discuss the molecular regulation of its localization. Remarkably, a number of LGN‐independent mechanisms were described recently, whereas NuMA remains central in most pathways involved in recruiting force generators at the cell cortex. We also describe the emerging role of the actin cortex in spindle orientation and discuss how dynamic astral microtubule formation is involved. We further give an overview on instructive external signals that control spindle orientation in tissues. Finally, we discuss the influence of cell geometry and mechanical forces on spindle orientation.

[1]  Shannon F. Stewman,et al.  The actin-binding ERM protein Moesin binds to and stabilizes microtubules at the cell cortex , 2013, The Journal of cell biology.

[2]  P. Insel,et al.  Identification and cDNA cloning of a novel human mosaic protein, LGN, based on interaction with G alpha i2. , 1996, Gene.

[3]  M. Gotta,et al.  Control of Embryonic Spindle Positioning and Gα Activity by C. elegans RIC-8 , 2004, Current Biology.

[4]  W. Bement,et al.  Myosin-10 and actin filaments are essential for mitotic spindle function , 2008, The Journal of cell biology.

[5]  H. Pasolli,et al.  Developmental roles for Srf, cortical cytoskeleton and cell shape in epidermal spindle orientation , 2011, Nature Cell Biology.

[6]  C. Heisenberg,et al.  Tension-oriented cell divisions limit anisotropic tissue tension in epithelial spreading during zebrafish epiboly , 2013, Nature Cell Biology.

[7]  I. Macara,et al.  Mammalian Pins Is a Conformational Switch that Links NuMA to Heterotrimeric G Proteins , 2004, Cell.

[8]  S. Akbarian,et al.  A Unique Set of Centrosome Proteins Requires Pericentrin for Spindle-Pole Localization and Spindle Orientation , 2014, Current Biology.

[9]  María Almonacid,et al.  Actin-based spindle positioning: new insights from female gametes , 2014, Journal of Cell Science.

[10]  Daniel A Fletcher,et al.  Cytoplasmic Volume Modulates Spindle Size During Embryogenesis , 2013, Science.

[11]  A. Bauer,et al.  Regulation of Notch signalling by non-visual β-arrestin , 2005, Nature Cell Biology.

[12]  H. Gröne,et al.  Semaphorin-Plexin Signaling Controls Mitotic Spindle Orientation during Epithelial Morphogenesis and Repair. , 2015, Developmental cell.

[13]  Yohanns Bellaïche,et al.  Drosophila Ric-8 regulates Gαi cortical localization to promote Gαi-dependent planar orientation of the mitotic spindle during asymmetric cell division , 2005, Nature Cell Biology.

[14]  C. Ponting,et al.  MISP is a novel Plk1 substrate required for proper spindle orientation and mitotic progression , 2013, The Journal of cell biology.

[15]  Y. Bellaïche,et al.  The Fz-Dsh planar cell polarity pathway induces oriented cell division via Mud/NuMA in Drosophila and zebrafish. , 2010, Developmental cell.

[16]  C. A. Johnston,et al.  Warts Phosphorylates Mud to Promote Pins-Mediated Mitotic Spindle Orientation in Drosophila, Independent of Yorkie , 2015, Current Biology.

[17]  Wei-Lih Lee,et al.  Astral microtubule asymmetry provides directional cues for spindle positioning in budding yeast. , 2012, Experimental cell research.

[18]  M. Glotzer,et al.  The 3Ms of central spindle assembly: microtubules, motors and MAPs , 2009, Nature Reviews Molecular Cell Biology.

[19]  C. Doe,et al.  Canoe binds RanGTP to promote PinsTPR/Mud-mediated spindle orientation , 2011, The Journal of cell biology.

[20]  D. Drubin,et al.  Dynein-driven mitotic spindle positioning restricted to anaphase by She1p inhibition of dynactin recruitment. , 2009, Molecular biology of the cell.

[21]  Xavier Morin,et al.  Control of planar divisions by the G-protein regulator LGN maintains progenitors in the chick neuroepithelium , 2007, Nature Neuroscience.

[22]  P. Gönczy,et al.  NuMA interacts with phosphoinositides and links the mitotic spindle with the plasma membrane , 2014, The EMBO journal.

[23]  A. Muroyama,et al.  NuMA-microtubule interactions are critical for spindle orientation and the morphogenesis of diverse epidermal structures , 2016, eLife.

[24]  C. Doe,et al.  Formin-mediated actin polymerization cooperates with Mushroom body defect (Mud)–Dynein during Frizzled–Dishevelled spindle orientation , 2013, Journal of Cell Science.

[25]  E. Nishida,et al.  Integrin‐mediated adhesion orients the spindle parallel to the substratum in an EB1‐ and myosin X‐dependent manner , 2007, The EMBO journal.

[26]  David P. Siderovski,et al.  Ric-8 controls Drosophila neural progenitor asymmetric division by regulating heterotrimeric G proteins , 2005, Nature Cell Biology.

[27]  Yohanns Bellaiche,et al.  Huntingtin Is Required for Mitotic Spindle Orientation and Mammalian Neurogenesis , 2010, Neuron.

[28]  J. Labbé,et al.  Heterotrimeric G protein signaling functions with dynein to promote spindle positioning in C. elegans , 2007, The Journal of cell biology.

[29]  Wieland B Huttner,et al.  Aspm specifically maintains symmetric proliferative divisions of neuroepithelial cells , 2006, Proceedings of the National Academy of Sciences.

[30]  X. Morin,et al.  Dlg1 controls planar spindle orientation in the neuroepithelium through direct interaction with LGN , 2014, The Journal of cell biology.

[31]  C. Doe,et al.  Microtubule-Induced Pins/Gαi Cortical Polarity in Drosophila Neuroblasts , 2005, Cell.

[32]  N. Fuse,et al.  Differential functions of G protein and Baz–aPKC signaling pathways in Drosophila neuroblast asymmetric division , 2004, The Journal of cell biology.

[33]  Manfred Gessler,et al.  Loss of Fat4 disrupts PCP signaling and oriented cell division and leads to cystic kidney disease , 2008, Nature Genetics.

[34]  T. Miki,et al.  Ric-8A and Giα Recruit LGN, NuMA, and Dynein to the Cell Cortex To Help Orient the Mitotic Spindle , 2010, Molecular and Cellular Biology.

[35]  M. Roldán,et al.  Spindle alignment is achieved without rotation after the first cell cycle in Drosophila embryonic neuroblasts , 2009, Development.

[36]  S. Bowman,et al.  Drosophila Ric-8 is essential for plasma-membrane localization of heterotrimeric G proteins , 2005, Nature Cell Biology.

[37]  S. Yoshiura,et al.  Tre1 GPCR signaling orients stem cell divisions in the Drosophila central nervous system. , 2012, Developmental cell.

[38]  P. Gönczy,et al.  RIC-8 Is Required for GPR-1/2-Dependent Gα Function during Asymmetric Division of C. elegans Embryos , 2004, Cell.

[39]  C. Doe,et al.  Identification of an Aurora-A/PinsLINKER/ Dlg Spindle Orientation Pathway using Induced Cell Polarity in S2 Cells , 2009, Cell.

[40]  F. Perez,et al.  γ‐Tubulin Ring Complexes and EB1 play antagonistic roles in microtubule dynamics and spindle positioning , 2014, The EMBO journal.

[41]  Wei-Lih Lee,et al.  Regulated offloading of cytoplasmic dynein from microtubule plus ends to the cortex. , 2011, Developmental cell.

[42]  F. McNally,et al.  Mechanisms of spindle positioning , 2013, The Journal of cell biology.

[43]  J. Knoblich,et al.  The PDZ Protein Canoe Regulates the Asymmetric Division of Drosophila Neuroblasts and Muscle Progenitors , 2008, Current Biology.

[44]  K. Ramyar,et al.  A Complex of NuMA and Cytoplasmic Dynein Is Essential for Mitotic Spindle Assembly , 1996, Cell.

[45]  Michel Bornens,et al.  External forces control mitotic spindle positioning , 2011, Nature Cell Biology.

[46]  C. Doe,et al.  Structure of an Enzyme-Derived Phosphoprotein Recognition Domain , 2012, PloS one.

[47]  Andrew E. Pelling,et al.  Moesin Controls Cortical Rigidity, Cell Rounding, and Spindle Morphogenesis during Mitosis , 2008, Current Biology.

[48]  Hans Clevers,et al.  Spindle orientation bias in gut epithelial stem cell compartments is lost in precancerous tissue. , 2010, Cell stem cell.

[49]  S. Grill,et al.  RGS-7 Completes a Receptor-Independent Heterotrimeric G Protein Cycle to Asymmetrically Regulate Mitotic Spindle Positioning in C. elegans , 2004, Cell.

[50]  B. Baum,et al.  Shaping up to divide: coordinating actin and microtubule cytoskeletal remodelling during mitosis. , 2014, Seminars in cell & developmental biology.

[51]  S. McKinney,et al.  Epithelial junctions maintain tissue architecture by directing planar spindle orientation , 2013, Nature.

[52]  E. Fuchs,et al.  Par3–mInsc and Gαi3 cooperate to promote oriented epidermal cell divisions through LGN , 2014, Nature Cell Biology.

[53]  J. Ahringer,et al.  Distinct roles for Galpha and Gbetagamma in regulating spindle position and orientation in Caenorhabditis elegans embryos. , 2001, Nature cell biology.

[54]  Revolving movement of a dynamic cluster of actin filaments during mitosis , 2010, The Journal of cell biology.

[55]  J. Cooper,et al.  Function of dynein in budding yeast: mitotic spindle positioning in a polarized cell. , 2009, Cell motility and the cytoskeleton.

[56]  A. Bretscher,et al.  Organizing the cell cortex: the role of ERM proteins , 2010, Nature Reviews Molecular Cell Biology.

[57]  M. Bownes,et al.  Locomotion defects, together with Pins, regulates heterotrimeric G-protein signaling during Drosophila neuroblast asymmetric divisions. , 2005, Genes & development.

[58]  D. St Johnston,et al.  Spindle orientation: What if it goes wrong? , 2014, Seminars in cell & developmental biology.

[59]  P. Bryant,et al.  The Partner of Inscuteable/Discs-Large Complex Is Required to Establish Planar Polarity during Asymmetric Cell Division in Drosophila , 2001, Cell.

[60]  Jan Ellenberg,et al.  The transition from meiotic to mitotic spindle assembly is gradual during early mammalian development , 2012, The Journal of cell biology.

[61]  T. Raabe,et al.  Drosophila Pins-binding protein Mud regulates spindle-polarity coupling and centrosome organization , 2006, Nature Cell Biology.

[62]  R. Petralia,et al.  mPins modulates PSD-95 and SAP102 trafficking and influences NMDA receptor surface expression , 2005, Nature Cell Biology.

[63]  Y. Bellaïche,et al.  Cell division orientation and planar cell polarity pathways. , 2009, Seminars in cell & developmental biology.

[64]  M. Mapelli,et al.  NuMA Phosphorylation by Aurora-A Orchestrates Spindle Orientation , 2016, Current Biology.

[65]  P. Gönczy,et al.  NuMA phosphorylation dictates dynein-dependent spindle positioning , 2013, Cell cycle.

[66]  M. Fuller,et al.  Orientation of Asymmetric Stem Cell Division by the APC Tumor Suppressor and Centrosome , 2003, Science.

[67]  Nicolas Minc,et al.  Influence of Cell Geometry on Division-Plane Positioning , 2011, Cell.

[68]  O. Hertwig,et al.  Das Problem der Befruchtung und der Isotropie des Eies, eine Theorie der Vererbung , 1884 .

[69]  Taryn E. Gillies,et al.  Cell Division Orientation in Animals , 2011, Current Biology.

[70]  A. Merdes,et al.  A lateral belt of cortical LGN and NuMA guides mitotic spindle movements and planar division in neuroepithelial cells , 2011, The Journal of cell biology.

[71]  S. Doxsey,et al.  Rab11 endosomes contribute to mitotic spindle organization and orientation. , 2014, Developmental cell.

[72]  S. van den Heuvel,et al.  A complex of LIN-5 and GPR proteins regulates G protein signaling and spindle function in C elegans. , 2003, Genes & development.

[73]  A. Merdes,et al.  Direct binding of NuMA to tubulin is mediated by a novel sequence motif in the tail domain that bundles and stabilizes microtubules. , 2002, Journal of cell science.

[74]  D. St Johnston,et al.  Discs Large Links Spindle Orientation to Apical-Basal Polarity in Drosophila Epithelia , 2013, Current Biology.

[75]  E. Betzig,et al.  A Localized Wnt Signal Orients Asymmetric Stem Cell Division in Vitro , 2013, Science.

[76]  Wei-Lih Lee,et al.  Microtubule-dependent path to the cell cortex for cytoplasmic dynein in mitotic spindle orientation , 2011, Bioarchitecture.

[77]  D. Morton,et al.  Identification of genes required for cytoplasmic localization in early C. elegans embryos , 1988, Cell.

[78]  Andrea Dimitracopoulos,et al.  Mitotic rounding alters cell geometry to ensure efficient bipolar spindle formation. , 2013, Developmental cell.

[79]  S. Fraser,et al.  Stereotypical Cell Division Orientation Controls Neural Rod Midline Formation in Zebrafish , 2010, Current Biology.

[80]  C. Doe,et al.  The NuMA-related Mud protein binds Pins and regulates spindle orientation in Drosophila neuroblasts , 2006, Nature Cell Biology.

[81]  Y. Wang,et al.  Mammalian spindle orientation and position respond to changes in cell shape in a dynein-dependent fashion. , 2000, Molecular biology of the cell.

[82]  Hiroshi Kiyonari,et al.  Neuroepithelial progenitors undergo LGN-dependent planar divisions to maintain self-renewability during mammalian neurogenesis , 2008, Nature Cell Biology.

[83]  Andrew R. Harris,et al.  Emergence of homeostatic epithelial packing and stress dissipation through divisions oriented along the long cell axis , 2015, Proceedings of the National Academy of Sciences.

[84]  P. Gönczy,et al.  Cortical dynein is critical for proper spindle positioning in human cells , 2012, The Journal of cell biology.

[85]  J. Ahringer,et al.  Asymmetrically Distributed C. elegans Homologs of AGS3/PINS Control Spindle Position in the Early Embryo , 2003, Current Biology.

[86]  Manuel Théry,et al.  Experimental and theoretical study of mitotic spindle orientation , 2007, Nature.

[87]  J. Knoblich,et al.  Heterotrimeric G Proteins Direct Two Modes of Asymmetric Cell Division in the Drosophila Nervous System , 2001, Cell.

[88]  C. Doe,et al.  The tumour-suppressor genes lgl and dlg regulate basal protein targeting in Drosophila neuroblasts , 2000, Nature.

[89]  K. Kemphues,et al.  Control of cleavage spindle orientation in Caenorhabditis elegans: the role of the genes par-2 and par-3. , 1995, Genetics.

[90]  R. Medema,et al.  Chromosome misalignments induce spindle‐positioning defects , 2016, EMBO reports.

[91]  P. Gönczy,et al.  Coupling of cortical dynein and Gα proteins mediates spindle positioning in Caenorhabditis elegans , 2007, Nature Cell Biology.

[92]  Lennart Kester,et al.  Differential proliferation rates generate patterns of mechanical tension that orient tissue growth , 2013, The EMBO journal.

[93]  X. Morin,et al.  An oblique view on the role of spindle orientation in vertebrate neurogenesis , 2012, Development, growth & differentiation.

[94]  J. Januschke,et al.  Stem cell decisions: A twist of fate or a niche market? , 2014, Seminars in cell & developmental biology.

[95]  Thomas Lecuit,et al.  A global pattern of mechanical stress polarizes cell divisions and cell shape in the growing Drosophila wing disc , 2013, Development.

[96]  S. Bowman,et al.  The Drosophila NuMA Homolog Mud regulates spindle orientation in asymmetric cell division. , 2006, Developmental cell.

[97]  E. V. van Munster,et al.  Conformational changes in CLIP-170 regulate its binding to microtubules and dynactin localization , 2004, The Journal of cell biology.

[98]  M. Gotta,et al.  Control of embryonic spindle positioning and Galpha activity by C. elegans RIC-8. , 2004, Current biology : CB.

[99]  Ralph Weissleder,et al.  WNT5A/JNK and FGF/MAPK Pathways Regulate the Cellular Events Shaping the Vertebrate Limb Bud , 2010, Current Biology.

[100]  S. Dedhar,et al.  Integrin-Linked Kinase links Dynactin-1/Dynactin-2 with cortical Integrin receptors to orient the mitotic spindle relative to the substratum , 2015, Scientific Reports.

[101]  Y. Bellaïche,et al.  Mitotic spindle orientation in asymmetric and symmetric cell divisions during animal development. , 2011, Developmental cell.

[102]  J. Ross,et al.  MAP4 and CLASP1 operate as a safety mechanism to maintain a stable spindle position in mitosis , 2011, Nature Cell Biology.

[103]  Tomoyuki U. Tanaka.  Kinetochore–microtubule interactions: steps towards bi‐orientation , 2010, The EMBO journal.

[104]  I. Macara,et al.  A mammalian Partner of inscuteable binds NuMA and regulates mitotic spindle organization , 2001, Nature Cell Biology.

[105]  H. Pasolli,et al.  Asymmetric Cell Divisions Promote Notch-Dependent Epidermal Differentiation , 2011, Nature.

[106]  Matthieu Piel,et al.  Exploring the function of cell shape and size during mitosis. , 2014, Developmental cell.

[107]  M. Mapelli,et al.  Concomitant binding of Afadin to LGN and F-actin directs planar spindle orientation , 2015, Nature Structural &Molecular Biology.

[108]  Jing Liu,et al.  LGN regulates mitotic spindle orientation during epithelial morphogenesis , 2010, The Journal of cell biology.

[109]  S. Humbert,et al.  Huntingtin Regulates Mammary Stem Cell Division and Differentiation , 2014, Stem cell reports.

[110]  C. Doe,et al.  Extrinsic cues orient the cell division axis in Drosophila embryonic neuroblasts , 2006, Development.

[111]  Scott E. Fraser,et al.  Planar cell polarity signalling controls cell division orientation during zebrafish gastrulation , 2004, Nature.

[112]  P. Meraldi,et al.  Mitotic spindle (DIS)orientation and DISease: Cause or consequence? , 2012, The Journal of cell biology.

[113]  Elaine Fuchs,et al.  Asymmetric cell divisions promote stratification and differentiation of mammalian skin , 2005, Nature.

[114]  E. Knust,et al.  Drosophila Atypical Protein Kinase C Associates with Bazooka and Controls Polarity of Epithelia and Neuroblasts , 2000, The Journal of cell biology.

[115]  A. Pierani,et al.  Cerebrospinal fluid-derived Semaphorin3B orients neuroepithelial cell divisions in the apicobasal axis , 2015, Nature Communications.

[116]  C. Heisenberg,et al.  Anthrax toxin receptor 2a controls mitotic spindle positioning , 2012, Nature Cell Biology.

[117]  X. Morin,et al.  Analysis of partner of inscuteable, a Novel Player of Drosophila Asymmetric Divisions, Reveals Two Distinct Steps in Inscuteable Apical Localization , 2000, Cell.

[118]  J. Shabanowitz,et al.  Par3 Controls Epithelial Spindle Orientation by aPKC-Mediated Phosphorylation of Apical Pins , 2010, Current Biology.

[119]  Daniel J. Muller,et al.  Hydrostatic pressure and the actomyosin cortex drive mitotic cell rounding , 2011, Nature.

[120]  W. Huttner,et al.  Specific polar subpopulations of astral microtubules control spindle orientation and symmetric neural stem cell division , 2014, eLife.

[121]  I. Cheeseman,et al.  Cortical Dynein and Asymmetric Membrane Elongation Coordinately Position the Spindle in Anaphase , 2013, Cell.

[122]  Kai Dierkes,et al.  Monitoring actin cortex thickness in live cells. , 2013, Biophysical journal.

[123]  Yohanns Bellaïche,et al.  Drosophila Ric-8 regulates Galphai cortical localization to promote Galphai-dependent planar orientation of the mitotic spindle during asymmetric cell division. , 2005, Nature cell biology.

[124]  E. Nishida,et al.  PtdIns(3,4,5)P3 regulates spindle orientation in adherent cells. , 2007, Developmental cell.

[125]  David K. Lubensky,et al.  Epithelial tricellular junctions act as interphase cell shape sensors to orient mitosis , 2016, Nature.

[126]  Yanlan Mao,et al.  Planar polarization of the atypical myosin Dachs orients cell divisions in Drosophila. , 2011, Genes & development.

[127]  F. Matsuzaki,et al.  Role of cortical tumour-suppressor proteins in asymmetric division of Drosophila neuroblast , 2000, Nature.

[128]  I. Cheeseman,et al.  Chromosome and spindle pole-derived signals generate an intrinsic code for spindle position and orientation , 2012, Nature Cell Biology.

[129]  Manuel Théry,et al.  The extracellular matrix guides the orientation of the cell division axis , 2005, Nature Cell Biology.

[130]  A. Shevchenko,et al.  A protein complex containing Inscuteable and the Gα-binding protein Pins orients asymmetric cell divisions in Drosophila , 2000, Current Biology.

[131]  Mingjie Zhang,et al.  Guanylate kinase domains of the MAGUK family scaffold proteins as specific phospho‐protein‐binding modules , 2011, The EMBO journal.

[132]  P. Gönczy,et al.  NuMA phosphorylation by CDK1 couples mitotic progression with cortical dynein function , 2013, The EMBO journal.

[133]  Y. Jan,et al.  Role of inscuteable in orienting asymmetric cell divisions in Drosophila , 1996, Nature.

[134]  Antonio Baonza,et al.  The Orientation of Cell Divisions Determines the Shape of Drosophila Organs , 2005, Current Biology.

[135]  G. Danuser,et al.  Direct Microtubule-Binding by Myosin-10 Orients Centrosomes toward Retraction Fibers and Subcortical Actin Clouds. , 2015, Developmental cell.

[136]  Frank Jülicher,et al.  Cell Flow Reorients the Axis of Planar Polarity in the Wing Epithelium of Drosophila , 2010, Cell.

[137]  J. Ahringer,et al.  Distinct roles for Gα and Gβγ in regulating spindle position and orientation in Caenorhabditis elegans embryos , 2001, Nature Cell Biology.

[138]  A. Hyman,et al.  Membrane Invaginations Reveal Cortical Sites that Pull on Mitotic Spindles in One-Cell C. elegans Embryos , 2010, PloS one.

[139]  Q. Du,et al.  Evidence for dynein and astral microtubule–mediated cortical release and transport of Gαi/LGN/NuMA complex in mitotic cells , 2013, Molecular biology of the cell.

[140]  A. Echard,et al.  Moesin and its activating kinase Slik are required for cortical stability and microtubule organization in mitotic cells , 2008, The Journal of cell biology.

[141]  I. Ispolatov,et al.  Cell shape impacts on the positioning of the mitotic spindle with respect to the substratum , 2015, Molecular biology of the cell.

[142]  R. Medema,et al.  Astral microtubules control redistribution of dynein at the cell cortex to facilitate spindle positioning , 2014, Cell cycle.

[143]  D. Siderovski,et al.  Return of the GDI: the GoLoco motif in cell division. , 2004, Annual review of biochemistry.