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Published 5 July 2004. doi:10.1083/jcb.200402052
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 166, Number 1, 49-60
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Article

Mutations in orbit/mast reveal that the central spindle is comprised of two microtubule populations, those that initiate cleavage and those that propagate furrow ingression



Yoshihiro H. Inoue1, Matthew S. Savoian2, Takao Suzuki1, Endre Máthé2, Masa-Toshi Yamamoto1, and David M. Glover2

1 Drosophila Genetic Resource Center, Kyoto Institute of Technology, Kyoto 616-8354, Japan
2 Cancer Research UK Cell Cycle Genetics Research Group, Department of Genetics, University of Cambridge, Cambridge CB2 3EH, England, UK

Address correspondence to Y.H. Inoue, Drosophila Genetic Resource Center, Kyoto Institute of Technology, Sagaippongi-cho, Ukyou-ku, Kyoto 616-8354, Japan. Tel.: 81-75-873-2653. Fax: 81-75-861-0881. email: yhinoue{at}ipc.kit.ac.jp


Abstract

We address the relative roles of astral and central spindle microtubules (MTs) in cytokinesis of Drosophila melanogaster primary spermatocytes. Time-lapse imaging studies reveal that the central spindle is comprised of two MT populations, "interior" central spindle MTs found within the spindle envelope and "peripheral" astral MTs that probe the cytoplasm and initiate cleavage furrows where they contact the cortex and form overlapping bundles. The MT-associated protein Orbit/Mast/CLASP concentrates on interior rather than peripheral central spindle MTs. Interior MTs are preferentially affected in hypomorphic orbit mutants, and consequently the interior central spindle fails to form or is unstable. In contrast, peripheral MTs still probe the cortex and form regions of overlap that recruit the Pav-KLP motor and Aurora B kinase. orbit mutants have disorganized or incomplete anillin and actin rings, and although cleavage furrows initiate, they ultimately regress. Our work identifies a new function for Orbit/Mast/CLASP and identifies a novel MT population involved in cleavage furrow initiation.

Key Words: cytokinesis; Drosophila; meiosis; MAP; CLASP

Y.H. Inoue and M.S. Savoian contributed equally to this paper.

The online version of this article includes supplemental material.

Abbreviations used in this paper: DIC, differential interference contrast; MAP, MT-associated protein; MT, microtubule.


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