Physical Description of Mitotic Spindle Orientation During Cell Division

Andrea Jiménez-Dalmaroni; Manuel Théry; Victor Racine; Michel Bornens; Frank Jülicher

doi:10.1016/j.bpj.2008.12.1049

Plk1 regulates spindle orientation by phosphorylating NuMA in human cells

Life Science Alliance ◽

10.26508/lsa.201800223 ◽

2018 ◽

Vol 1 (6) ◽

pp. e201800223 ◽

Cited By ~ 14

Author(s):

Shrividya Sana ◽

Riya Keshri ◽

Ashwathi Rajeevan ◽

Sukriti Kapoor ◽

Sachin Kotak

Keyword(s):

Cell Division ◽

Mitotic Spindle ◽

Molecular Mechanisms ◽

Spindle Pole ◽

Spindle Orientation ◽

Cell Cortex ◽

Division Plane ◽

Evolutionarily Conserved ◽

Cortical Localization ◽

Proper Orientation

Proper orientation of the mitotic spindle defines the correct division plane and is essential for accurate cell division and development. In metazoans, an evolutionarily conserved complex comprising of NuMA/LGN/Gαi regulates proper orientation of the mitotic spindle by orchestrating cortical dynein levels during metaphase. However, the molecular mechanisms that modulate the spatiotemporal dynamics of this complex during mitosis remain elusive. Here, we report that acute inactivation of Polo-like kinase 1 (Plk1) during metaphase enriches cortical levels of dynein/NuMA/LGN and thus influences spindle orientation. We establish that this impact of Plk1 on cortical levels of dynein/NuMA/LGN is through NuMA, but not via dynein/LGN. Moreover, we reveal that Plk1 inhibition alters the dynamic behavior of NuMA at the cell cortex. We further show that Plk1 directly interacts and phosphorylates NuMA. Notably, NuMA-phosphorylation by Plk1 impacts its cortical localization, and this is needed for precise spindle orientation during metaphase. Overall, our finding connects spindle-pole pool of Plk1 with cortical NuMA and answers a long-standing puzzle about how spindle-pole Plk1 gradient dictates proper spindle orientation for error-free mitosis.

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Robust control of mitotic spindle orientation in the developing epidermis

The Journal of Cell Biology ◽

10.1083/jcb.201008001 ◽

2010 ◽

Vol 191 (5) ◽

pp. 915-922 ◽

Cited By ~ 106

Author(s):

Nicholas D. Poulson ◽

Terry Lechler

Keyword(s):

Cell Division ◽

Robust Control ◽

Progenitor Cells ◽

Mitotic Spindle ◽

Asymmetric Cell Division ◽

Transcriptional Regulator ◽

Spindle Orientation ◽

Control Points ◽

External Cues ◽

Increase Surface Area

Progenitor cells must balance self-amplification and production of differentiated progeny during development and homeostasis. In the epidermis, progenitors divide symmetrically to increase surface area and asymmetrically to promote stratification. In this study, we show that individual epidermal cells can undergo both types of division, and therefore, the balance is provided by the sum of individual cells’ choices. In addition, we define two control points for determining a cell’s mode of division. First is the expression of the mouse Inscuteable gene, which is sufficient to drive asymmetric cell division (ACD). However, there is robust control of division orientation as excessive ACDs are prevented by a change in the localization of NuMA, an effector of spindle orientation. Finally, we show that p63, a transcriptional regulator of stratification, does not control either of these processes. These data have uncovered two important regulatory points controlling ACD in the epidermis and allow a framework for analysis of how external cues control this important choice.

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Cell contacts orient some cell division axes in the Caenorhabditis elegans embryo.

The Journal of Cell Biology ◽

10.1083/jcb.129.4.1071 ◽

1995 ◽

Vol 129 (4) ◽

pp. 1071-1080 ◽

Cited By ~ 79

Author(s):

B Goldstein

Keyword(s):

Cell Division ◽

Caenorhabditis Elegans ◽

Mitotic Spindle ◽

Spindle Orientation ◽

Cell Contact ◽

Specific Cell ◽

Cell Contacts ◽

Established Cell ◽

A Site ◽

Cell Cell

Cells of the early Caenorhabditis elegans embryo divide in an invariant pattern. Here I show that the division axes of some early cells (EMS and E) are controlled by specific cell-cell contacts (EMS-P2 or E-P3 contact). Altering the orientation of contact between these cells alters the axis along which the mitotic spindle is established, and hence the orientation of cell division. Contact-dependent mitotic spindle orientation appears to work by establishing a site of the type described by Hyman and White (1987. J. Cell Biol. 105:2123-2135) in the cortex of the responding cell: one centrosome moves toward the site of cell-cell contact during centrosome rotation in both intact embryos and reoriented cell pairs. The effect is especially apparent when two donor cells are placed on one side of the responding cell: both centrosomes are "captured," pulling the nucleus to one side of the cell. No centrosome rotation occurs in the absence of cell-cell contact, nor in nocodazole-treated cell pairs. The results suggest that some of the cortical sites described by Hyman and White are established cell autonomously (in P1, P2, and P3), and some are established by cell-cell contact (in EMS and E). Additional evidence presented here suggests that in the EMS cell, contact-dependent spindle orientation ensures a cleavage plane that will partition developmental information, received by induction, to one of EMS's daughter cells.

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Microtubule end tethering of a processive kinesin-8 motor Kif18b is required for spindle positioning

The Journal of Cell Biology ◽

10.1083/jcb.201705209 ◽

2018 ◽

Vol 217 (7) ◽

pp. 2403-2416 ◽

Cited By ~ 18

Author(s):

Toni McHugh ◽

Agata A. Gluszek ◽

Julie P.I. Welburn

Keyword(s):

Cell Division ◽

Mitotic Spindle ◽

Essential Role ◽

Gene Knockout ◽

Spindle Orientation ◽

Binding Region ◽

Spindle Positioning ◽

Astral Microtubule ◽

In Vitro Reconstitution

Mitotic spindle positioning specifies the plane of cell division during anaphase. Spindle orientation and positioning are therefore critical to ensure symmetric division in mitosis and asymmetric division during development. The control of astral microtubule length plays an essential role in positioning the spindle. In this study, using gene knockout, we show that the kinesin-8 Kif18b controls microtubule length to center the mitotic spindle at metaphase. Using in vitro reconstitution, we reveal that Kif18b is a highly processive plus end–directed motor that uses a C-terminal nonmotor microtubule-binding region to accumulate at growing microtubule plus ends. This region is regulated by phosphorylation to spatially control Kif18b accumulation at plus ends and is essential for Kif18b-dependent spindle positioning and regulation of microtubule length. Finally, we demonstrate that Kif18b shortens microtubules by increasing the catastrophe rate of dynamic microtubules. Overall, our work reveals that Kif18b uses its motile properties to reach microtubule ends, where it regulates astral microtubule length to ensure spindle centering.

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Microtubule end tethering of a processive Kinesin-8 motor Kif18b is required for spindle positioning

10.1101/286658 ◽

2018 ◽

Cited By ~ 2

Author(s):

Toni McHugh ◽

Agata Gluszek-Kustusz ◽

Julie P.I. Welburn

Keyword(s):

Cell Division ◽

Mitotic Spindle ◽

Essential Role ◽

Gene Knockout ◽

Integrated Approach ◽

Spindle Orientation ◽

Binding Region ◽

Spindle Positioning ◽

Symmetric Division ◽

Astral Microtubule

AbstractMitotic spindle positioning specifies the plane of cell division during anaphase. Spindle orientation and positioning is therefore critical to ensure symmetric division in mitosis and asymmetric division during development. The control of astral microtubule length plays an essential role in positioning the spindle. Here we show using gene knockout that the Kinesin-8 Kif18b controls microtubule length to center the mitotic spindle at metaphase. Using an integrated approach, we reveal that Kif18b is a highly processive plus end-directed motor that uses a C-terminal non-motor microtubule-binding region to accumulate at growing microtubule plus ends. This region is regulated by phosphorylation to spatially control Kif18b accumulation at plus ends and is essential for Kif18b-dependent spindle positioning and regulation of microtubule length. Finally, we demonstrate that Kif18b shortens microtubules by increasing the catastrophe rate of dynamic microtubules. Overall, our work reveals that Kif18b utilizes its motile properties to reach microtubule ends where it regulates astral microtubule length to ensure spindle centering.

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Specific polar subpopulations of astral microtubules control spindle orientation and symmetric neural stem cell division

eLife ◽

10.7554/elife.02875 ◽

2014 ◽

Vol 3 ◽

Cited By ~ 33

Author(s):

Felipe Mora-Bermúdez ◽

Fumio Matsuzaki ◽

Wieland B Huttner

Keyword(s):

Cell Division ◽

Basal Cell ◽

Mitotic Spindle ◽

Asymmetric Cell Division ◽

Spindle Cell ◽

Spindle Orientation ◽

Cell Cortex ◽

Dividing Cells ◽

Stem Cell Division

Mitotic spindle orientation is crucial for symmetric vs asymmetric cell division and depends on astral microtubules. Here, we show that distinct subpopulations of astral microtubules exist, which have differential functions in regulating spindle orientation and division symmetry. Specifically, in polarized stem cells of developing mouse neocortex, astral microtubules reaching the apical and basal cell cortex, but not those reaching the central cell cortex, are more abundant in symmetrically than asymmetrically dividing cells and reduce spindle orientation variability. This promotes symmetric divisions by maintaining an apico-basal cleavage plane. The greater abundance of apical/basal astrals depends on a higher concentration, at the basal cell cortex, of LGN, a known spindle-cell cortex linker. Furthermore, newly developed specific microtubule perturbations that selectively decrease apical/basal astrals recapitulate the symmetric-to-asymmetric division switch and suffice to increase neurogenesis in vivo. Thus, our study identifies a novel link between cell polarity, astral microtubules, and spindle orientation in morphogenesis.

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Activation of Discs large by aPKC aligns the mitotic spindle to the polarity axis during asymmetric cell division

eLife ◽

10.7554/elife.32137 ◽

2017 ◽

Vol 6 ◽

Cited By ~ 4

Author(s):

Ognjen Golub ◽

Brett Wee ◽

Rhonda A Newman ◽

Nicole M Paterson ◽

Kenneth E Prehoda

Keyword(s):

Cell Division ◽

Mitotic Spindle ◽

Asymmetric Cell Division ◽

Intramolecular Interaction ◽

Spindle Orientation ◽

Orientation Factor ◽

Daughter Cells ◽

Kinase C ◽

Discs Large ◽

Fate Determinants

Asymmetric division generates cellular diversity by producing daughter cells with different fates. In animals, the mitotic spindle aligns with Par complex polarized fate determinants, ensuring that fate determinant cortical domains are bisected by the cleavage furrow. Here, we investigate the mechanisms that couple spindle orientation to polarity during asymmetric cell division of Drosophila neuroblasts. We find that the tumor suppressor Discs large (Dlg) links the Par complex component atypical Protein Kinase C (aPKC) to the essential spindle orientation factor GukHolder (GukH). Dlg is autoinhibited by an intramolecular interaction between its SH3 and GK domains, preventing Dlg interaction with GukH at cortical sites lacking aPKC. When co-localized with aPKC, Dlg is phosphorylated in its SH3 domain which disrupts autoinhibition and allows GukH recruitment by the GK domain. Our work establishes a molecular connection between the polarity and spindle orientation machineries during asymmetric cell division.

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