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Originally published as MBC in Press, 10.1091/mbc.E06-11-0994 on June 6, 2007

Vol. 18, Issue 8, 3094-3104, August 2007

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Poleward Tubulin Flux in Spindles: Regulation and Function in Mitotic CellsFormula Formula

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

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

Submitted November 8, 2006; Revised May 24, 2007; Accepted May 29, 2007
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 RNA interference 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- or down-regulated or restored after 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 establishment of normal spindle length during prometaphase/metaphase.

This was published online ahead of print in MBC in Press (http://www.molbiolcell.org/cgi/doi/10.1091/mbc.E06-11-0994) on June 6, 2007.

Formula Formula The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).

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

Abbreviations used: FRAP, fluorescence recovery after photobleaching.




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