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Drosophila Klp67A binds prophase kinetochores to subsequently regulate congression and spindle length

Savoian, Matthew S. and Glover, David M. (2010) Drosophila Klp67A binds prophase kinetochores to subsequently regulate congression and spindle length. Journal of Cell Science, 123 (5). pp. 767-776. ISSN 0021-9533. https://resolver.caltech.edu/CaltechAUTHORS:20200930-144713931

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[img] Image (JPEG) (Fig. S1. Klp67AEGFP variants used in this study) - Supplemental Material
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[img] Image (JPEG) (Fig. S2. Klp67AEGFP variant distributions during interphase) - Supplemental Material
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[img] Image (JPEG) (Supplemental Figure S3. Endogenous Klp67A labels the metaphase spindle and collects adjacent to the centromeres) - Supplemental Material
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[img] Image (JPEG) (Fig. S4. Klp67A depolymerises interphase MTs in a motor-activity-dependent manner) - Supplemental Material
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[img] Image (JPEG) (Fig. S5. Tail-only Klp67AEGFP localises with centrosomes in an MT-dependent manner) - Supplemental Material
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[img] Image (JPEG) (Fig. S6. Incubation with 25 µM colchicine depolymerises spindle-associated MTs, but does not prevent the association of Klp67A with kinetochores) - Supplemental Material
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[img] Video (QuickTime) (Movie 1. Prometaphase onset in a Klp67A(WT)EGFP-expressing neuroblast ) - Supplemental Material
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[img] Video (QuickTime) (Movie 2. Prometaphase onset in a Klp67A(RIG)EGFP-expressing neuroblast) - Supplemental Material
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[img] Video (QuickTime) (Movie 3. Prometaphase onset in a Klp67A(TL)EGFP-expressing neuroblast) - Supplemental Material
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Abstract

The kinesin-8 proteins are a family of microtubule-depolymerising motor molecules, which, despite their highly conserved roles in chromosome alignment and spindle dynamics, remain poorly characterised. Here, we report that the Drosophila kinesin-8 protein, Klp67A, exists in two spatially and functionally separable metaphase pools: at kinetochores and along the spindle. Fixed and live-cell analyses of different Klp67A recombinant variants indicate that this kinesin-8 first collects at kinetochores during prophase and, by metaphase, localises to the kinetochore outerplate. Although the catalytic motor activity of Klp67A is required for efficient kinetochore recruitment at all times, microtubules are entirely dispensable for this process. The tail of Klp67A does not play a role in kinetochore accumulation, but is both necessary and sufficient for spindle association. Using functional assays, we reveal that chromosome position and spindle length are determined by the microtubule-depolymerising motor activity of Klp67A exclusively when located at kinetochores, but not along the spindle. These data reveal that, unlike other metazoan kinesin-8 proteins, Klp67A binds the nascent prophase and mature metaphase kinetochore. From this location, Klp67A uses its motor activity to ensure chromosome alignment and proper spindle length.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1242/jcs.055905DOIArticle
http://jcs.biologists.org/lookup/suppl/doi:10.1242/jcs.055905/-/DC1PublisherSupporting Information
ORCID:
AuthorORCID
Savoian, Matthew S.0000-0002-2594-3879
Glover, David M.0000-0003-0956-0103
Additional Information:© 2010. Published by The Company of Biologists Ltd. Accepted November 30, 2009. Published online February 17, 2010. The authors wish to thank the members of the Glover and Segal laboratories for insightful conversations during this work. We further acknowledge Pier Paolo D'Avino and Feng Chen, University of Cambridge, UK for allowing us to use their modified pUAST vector before publication. This work was made possible by a CRUK program grant to D.M.G.
Funders:
Funding AgencyGrant Number
Cancer Research UKUNSPECIFIED
Subject Keywords:Kinesin-8, Mitosis, Microtubule, Depolymerase, Motor protein
Issue or Number:5
Record Number:CaltechAUTHORS:20200930-144713931
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20200930-144713931
Official Citation:Drosophila Klp67A binds prophase kinetochores to subsequently regulate congression and spindle length Matthew S. Savoian, David M. Glover Journal of Cell Science 2010 123: 767-776; doi: 10.1242/jcs.055905
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:105701
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:30 Sep 2020 22:47
Last Modified:30 Sep 2020 22:47

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