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MBC in Press, published online ahead of print June 6, 2007
Mol. Biol. Cell 10.1091/mbc.E06-11-0994

A more recent version of this article appeared on August 1, 2007
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Submitted on November 8, 2006
Revised on May 24, 2007
Accepted on May 29, 2007

Poleward Tubulin Flux in Spindles: Regulation and Function in Mitotic Cells

Daniel W. Buster, Dong Zhang, and David J. Sharp

Department of Physiology and Biophysics, Albert Einstein College of Medicine, Bronx, NY 10461

Monitoring Editor: Yixian Zheng

The poleward flux of tubulin subunits through spindle microtubules is a striking and conserved phenomenon whose function and molecular components remain poorly understood. To screen for novel components of the flux machinery, we utilized RNAi to deplete regulators of microtubule dynamics, individually and in various combinations, from S2 cells and examined the resulting impact on flux rate. This led to the identification of two previously unknown flux inhibitors, KLP59C and KLP67A, and a flux promoter, Mini-spindles. Furthermore, we find that flux rate is regulated by functional antagonism among microtubule stabilizers and destabilizers specifically at plus-ends. Finally, by examining mitosis on spindles in which flux has been up/down-regulated or restored following the codepletion of antagonistic flux regulators, we show that flux is an integral contributor to anaphase A but is not responsible for chromosome congression, interkinetochore tension, or the regulation of spindle length during prometaphase/metaphase.

Address correspondence to: David J. Sharp (dsharp{at}aecom.yu.edu)






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