A Novel Small-molecule Inhibitor Reveals a Possible Role of Kinesin-5 in Anastral Spindle-pole Assembly

Overview

Journal J Cell Sci

Specialty Cell Biology

Date 2008 Jun 19

PMID 18559893

Citations 32

Authors

Aaron C Groen

Daniel Needleman

Clifford Brangwynne

Christain Gradinaru

Brandon Fowler

Ralph Mazitschek

Timothy J Mitchison

Affiliations

Soon will be listed here.

Abstract

The tetrameric plus-end-directed motor, kinesin-5, is essential for bipolar spindle assembly. Small-molecule inhibitors of kinesin-5 have been important tools for investigating its function, and some are currently under evaluation as anti-cancer drugs. Most inhibitors reported to date are ;non-competitive' and bind to a specific site on the motor head, trapping the motor in an ADP-bound state in which it has a weak but non-zero affinity for microtubules. Here, we used a novel ATP-competitive inhibitor, FCPT, developed at Merck (USA). We found that it induced tight binding of kinesin-5 onto microtubules in vitro. Using Xenopus egg-extract spindles, we found that FCPT not only blocked poleward microtubule sliding but also selectively induced loss of microtubules at the poles of bipolar spindles (and not asters or monoasters). We also found that the spindle-pole proteins TPX2 and gamma-tubulin became redistributed to the spindle equator, suggesting that proper kinesin-5 function is required for pole assembly.

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