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doi:10.1083/jcb.200802189
The Journal of Cell Biology
The Rockefeller University Press, 0021-9525 $30.00
© Kim et al.
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CENP-E combines a slow, processive motor and a flexible coiled coil to produce an essential motile kinetochore tether



Yumi Kim1,2, John E. Heuser5, Clare M. Waterman6, and Don W. Cleveland1,3,4

1 Ludwig Institute for Cancer Research, 2 Division of Biological Sciences, 3 School of Medicine, and 4 Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093
5 Department of Cell Biology, School of Medicine, Washington University, St. Louis, MO 63110
6 National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892

Correspondence to Don W. Cleveland: dcleveland{at}ucsd.edu

The mitotic kinesin centromere protein E (CENP-E) is an essential kinetochore component that directly contributes to the capture and stabilization of spindle microtubules by kinetochores. Although reduction in CENP-E leads to high rates of whole chromosome missegregation, neither its properties as a microtubule-dependent motor nor how it contributes to the dynamic linkage between kinetochores and microtubules is known. Using single-molecule assays, we demonstrate that CENP-E is a very slow, highly processive motor that maintains microtubule attachment for long periods. Direct visualization of full-length Xenopus laevis CENP-E reveals a highly flexible 230-nm coiled coil separating its kinetochore-binding and motor domains. We also show that full-length CENP-E is a slow plus end–directed motor whose activity is essential for metaphase chromosome alignment. We propose that the highly processive microtubule-dependent motor activity of CENP-E serves to power chromosome congression and provides a flexible, motile tether linking kinetochores to dynamic spindle microtubules.

Abbreviations used in this paper: CENP-E, centromere protein E; GMPCPP, guanosine-5'-([a,b]-methyleno) triphosphate; MSD, mean square displacement; TIRF, total internal reflection fluorescence.


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