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    Length control of the metaphase spindle

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    Author's final draft (556Kb)
    Figures (293.4Kb)
    Klp67A-GFP Dynamics upon Colchicine Treatment (2.369Mb)
    Fluorescent Speckle Microscopy of GFP-Tubulin in Spindles without RNAi Treatment (2.057Mb)
    Fluorescent Speckle Microscopy of GFP-Tubulin in Spindles after Klp61F RNAi (6.348Mb)
    Date
    2005-09-30
    Author
    Goshima, Gohta  Concept link
    Wollman, Roy  Concept link
    Stuurman, Nico  Concept link
    Scholey, Jonathan M.  Concept link
    Vale, Ronald D.  Concept link
    Metadata
    Show full item record
    Citable URI
    https://hdl.handle.net/1912/291
    As published
    https://doi.org/10.1016/j.cub.2005.09.054
    Keyword
     RNAi; Mitosis; Microtubule; Drosophila S2 cell 
    Abstract
    The pole-to-pole distance of the metaphase spindle is reasonably constant in a given cell type; in the case of vertebrate female oocytes, this steady-state length can be maintained for substantial lengths of time, during which time microtubules remain highly dynamic. Although a number of molecular perturbations have been shown to influence spindle length, a global understanding of the factors that determine metaphase spindle length has not been achieved. Using the Drosophila S2 cell line, we depleted or overexpressed proteins that either generate sliding forces between spindle microtubules (Kinesin-5, Kinesin-14, dynein), promote microtubule polymerization (EB1, Mast/Orbit [CLASP], Minispindles [Dis1/XMAP215/TOG]) or depolymerization (Kinesin-8, Kinesin-13), or mediate sister-chromatid cohesion (Rad21) in order to explore how these forces influence spindle length. Using high-throughput automated microscopy and semiautomated image analyses of >4000 spindles, we found a reduction in spindle size after RNAi of microtubule-polymerizing factors or overexpression of Kinesin-8, whereas longer spindles resulted from the knockdown of Rad21, Kinesin-8, or Kinesin-13. In contrast, and differing from previous reports, bipolar spindle length is relatively insensitive to increases in motor-generated sliding forces. However, an ultrasensitive monopolar-to-bipolar transition in spindle architecture was observed at a critical concentration of the Kinesin-5 sliding motor. These observations could be explained by a quantitative model that proposes a coupling between microtubule depolymerization rates and microtubule sliding forces. By integrating extensive RNAi with high-throughput image-processing methodology and mathematical modeling, we reach to a conclusion that metaphase spindle length is sensitive to alterations in microtubule dynamics and sister-chromatid cohesion, but robust against alterations of microtubule sliding force.
    Description
    Author Posting. © The Authors, 2005. This is the author's version of the work. It is posted here by permission of Elsevier B. V. for personal use, not for redistribution. The definitive version was published in Current Biology 15 (2005): 1979-1988, doi:10.1016/j.cub.2005.09.054.
    Collections
    • Physiology
    Suggested Citation
    Preprint: Goshima, Gohta, Wollman, Roy, Stuurman, Nico, Scholey, Jonathan M., Vale, Ronald D., "Length control of the metaphase spindle", 2005-09-30, https://doi.org/10.1016/j.cub.2005.09.054, https://hdl.handle.net/1912/291
     
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