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Antagonistic activities of Klp10A and Orbit regulate spindle length, bipolarity and function in vivo

Laycock, Joseph E. and Savoian, Matthew S. and Glover, David M. (2006) Antagonistic activities of Klp10A and Orbit regulate spindle length, bipolarity and function in vivo. Journal of Cell Science, 119 (11). pp. 2354-2361. ISSN 0021-9533. https://resolver.caltech.edu/CaltechAUTHORS:20201002-140639625

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[img] Image (JPEG) (Fig. S1. S2 cell FACs profiles following RNAi of orbit and/or Klp59C, Klp59D and Klp67A) - Supplemental Material
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[img] Video (AVI) (Movie 1. Spindle formation in a control cell expressing GFP-tubulin) - Supplemental Material
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[img] Video (AVI) (Movie 2. Spindle formation in cells singly depleted of Klp10A or Orbit) - Supplemental Material
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[img] Video (AVI) (Movie 3. Dual RNAi of orbit and Klp10A prevents spindle collapse and rescues spindle bipolarity) - Supplemental Material
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Abstract

The metaphase-spindle steady-state length occurs as spindle microtubules `flux', incorporating new subunits at their plus ends, while simultaneously losing subunits from their minus ends. Orbit/Mast/CLASP is required for tubulin subunit addition at kinetochores, and several kinesins regulate spindle morphology and/or flux by serving as microtubule depolymerases. Here, we use RNA interference in S2 cells to examine the relationship between Orbit and the four predicted kinesin-type depolymerases encoded by the Drosophila genome (Klp10A, Klp59C, Klp59D and Klp67A). Single depletion of Orbit results in monopolar spindles, mitotic arrest and a subsequent increase in apoptotic cells. These phenotypes are rescued by co-depleting Klp10A but none of the other three depolymerases. Spindle bipolarity is restored by preventing the spindle collapse seen in cells that lack Orbit, leading to functional spindles that are similar to controls in shape and length. We conclude that Klp10A exclusively antagonises Orbit in the regulation of bipolar spindle formation and maintenance.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1242/jcs.02957DOIArticle
http://jcs.biologists.org/cgi/content/full/119/11/2354/DC1PublisherSupporting Information
ORCID:
AuthorORCID
Savoian, Matthew S.0000-0002-2594-3879
Glover, David M.0000-0003-0956-0103
Additional Information:© The Company of Biologists Limited 2006. Accepted February 23, 2006. Published online May 24, 2006. We thank E. Mathe for aid in the generating the Klp10A antibody. This work was made by possible grants to D.M.G. from the Cancer Research UK and Medical Research Council, and a studentship from the Biotechnology and Biological Sciences Research Council.
Funders:
Funding AgencyGrant Number
Cancer Research UKUNSPECIFIED
Medical Research Council (UK)UNSPECIFIED
Biotechnology and Biological Sciences Research Council (BBSRC)UNSPECIFIED
Subject Keywords:Kinesin, Mitosis, Microtubule, Catastrophe factor, Flux, CLASP
Issue or Number:11
Record Number:CaltechAUTHORS:20201002-140639625
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20201002-140639625
Official Citation:Antagonistic activities of Klp10A and Orbit regulate spindle length, bipolarity and function in vivo Joseph E. Laycock, Matthew S. Savoian, David M. Glover Journal of Cell Science 2006 119: 2354-2361; doi: 10.1242/jcs.02957
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:105759
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:05 Oct 2020 16:17
Last Modified:05 Oct 2020 16:17

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