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      Ect2/Pbl Acts via Rho and Polarity Proteins to Direct the Assembly of an Isotropic Actomyosin Cortex upon Mitotic Entry

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      1 , 2 , 1 , 1 , 1 ,
      Developmental Cell
      Cell Press

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          Summary

          Entry into mitosis is accompanied by profound changes in cortical actomyosin organization. Here, we delineate a pathway downstream of the RhoGEF Pbl/Ect2 that directs this process in a model epithelium. Our data suggest that the release of Pbl/Ect2 from the nucleus at mitotic entry drives Rho-dependent activation of Myosin-II and, in parallel, induces a switch from Arp2/3 to Diaphanous-mediated cortical actin nucleation that depends on Cdc42, aPKC, and Par6. At the same time, the mitotic relocalization of these apical protein complexes to more lateral cell surfaces enables Cdc42/aPKC/Par6 to take on a mitosis-specific function—aiding the assembly of a relatively isotropic metaphase cortex. Together, these data reveal how the repolarization and remodeling of the actomyosin cortex are coordinated upon entry into mitosis to provide cells with the isotropic and rigid form they need to undergo faithful chromosome segregation and division in a crowded tissue environment.

          Highlights

          • Pbl/Ect2 drives a shift in epithelial polarity upon entry into mitosis

          • Lateral spreading of Cdc42/aPKC/Par6 aids assembly of an isotropic metaphase cortex

          • Mitosis triggers a switch from Arp2/3 to Dia-mediated cortical actin nucleation

          Abstract

          Rosa et al. identify a pathway activated upon entry into mitosis that drives lateral spreading of “apical” polarity proteins to coordinate the remodeling of cell shape and cytoskeleton. This mitotic repurposing of polarity proteins generates the rigid rounded cortex required for successful division in a crowded tissue.

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          Most cited references62

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          Hydrostatic pressure and the actomyosin cortex drive mitotic cell rounding.

          During mitosis, adherent animal cells undergo a drastic shape change, from essentially flat to round. Mitotic cell rounding is thought to facilitate organization within the mitotic cell and be necessary for the geometric requirements of division. However, the forces that drive this shape change remain poorly understood in the presence of external impediments, such as a tissue environment. Here we use cantilevers to track cell rounding force and volume. We show that cells have an outward rounding force, which increases as cells enter mitosis. We find that this mitotic rounding force depends both on the actomyosin cytoskeleton and the cells' ability to regulate osmolarity. The rounding force itself is generated by an osmotic pressure. However, the actomyosin cortex is required to maintain this rounding force against external impediments. Instantaneous disruption of the actomyosin cortex leads to volume increase, and stimulation of actomyosin contraction leads to volume decrease. These results show that in cells, osmotic pressure is balanced by inwardly directed actomyosin cortex contraction. Thus, by locally modulating actomyosin-cortex-dependent surface tension and globally regulating osmotic pressure, cells can control their volume, shape and mechanical properties.
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            The extracellular matrix guides the orientation of the cell division axis.

            The cell division axis determines the future positions of daughter cells and is therefore critical for cell fate. The positioning of the division axis has been mostly studied in systems such as embryos or yeasts, in which cell shape is well defined. In these cases, cell shape anisotropy and cell polarity affect spindle orientation. It remains unclear whether cell geometry or cortical cues are determinants for spindle orientation in mammalian cultured cells. The cell environment is composed of an extracellular matrix (ECM), which is connected to the intracellular actin cytoskeleton via transmembrane proteins. We used micro-contact printing to control the spatial distribution of the ECM on the substrate and demonstrated that it has a role in determining the orientation of the division axis of HeLa cells. On the basis of our analysis of the average distributions of actin-binding proteins in interphase and mitosis, we propose that the ECM controls the location of actin dynamics at the membrane, and thus the segregation of cortical components in interphase. This segregation is further maintained on the cortex of mitotic cells and used for spindle orientation.
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              Cell polarity in eggs and epithelia: parallels and diversity.

              Cell polarity, the generation of cellular asymmetries, is necessary for diverse processes in animal cells, such as cell migration, asymmetric cell division, epithelial barrier function, and morphogenesis. Common mechanisms generate and transduce cell polarity in different cells, but cell type-specific processes are equally important. In this review, we highlight the similarities and differences between the polarity mechanisms in eggs and epithelia. We also highlight the prospects for future studies on how cortical polarity interfaces with other cellular processes, such as morphogenesis, exocytosis, and lipid signaling, and how defects in polarity contribute to tumor formation. Copyright 2010 Elsevier Inc. All rights reserved.
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                Author and article information

                Contributors
                Journal
                Dev Cell
                Dev. Cell
                Developmental Cell
                Cell Press
                1534-5807
                1878-1551
                09 March 2015
                09 March 2015
                : 32
                : 5
                : 604-616
                Affiliations
                [1 ]MRC Laboratory of Molecular Cell Biology, UCL, Gower Street, London WC1E 6BT, UK
                [2 ]Graduate Program in Areas of Basic and Applied Biology (GABBA), University of Porto, 4200-465 Porto, Portugal
                Author notes
                []Corresponding author b.baum@ 123456ucl.ac.uk
                Article
                S1534-5807(15)00035-0
                10.1016/j.devcel.2015.01.012
                4359025
                25703349
                c9799508-75c1-433d-a71b-60ee7108a8d5
                © 2015 The Authors

                This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

                History
                : 1 March 2014
                : 6 October 2014
                : 14 January 2015
                Categories
                Article

                Developmental biology
                Developmental biology

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