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Published online
doi:10.1083/jcb.200810091
The Journal of Cell Biology, Vol. 184, No. 5, 677-690
The Rockefeller University Press, 0021-9525 $30.00
© Silk et al.
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Article

Requirements for NuMA in maintenance and establishment of mammalian spindle poles



Alain D. Silk1,2, Andrew J. Holland1,2, and Don W. Cleveland1,2

1 Ludwig Institute for Cancer Research and 2 Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093

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

Microtubules of the mitotic spindle in mammalian somatic cells are focused at spindle poles, a process thought to include direct capture by astral microtubules of kinetochores and/or noncentrosomally nucleated microtubule bundles. By construction and analysis of a conditional loss of mitotic function allele of the nuclear mitotic apparatus (NuMA) protein in mice and cultured primary cells, we demonstrate that NuMA is an essential mitotic component with distinct contributions to the establishment and maintenance of focused spindle poles. When mitotic NuMA function is disrupted, centrosomes provide initial focusing activity, but continued centrosome attachment to spindle fibers under tension is defective, and the maintenance of focused kinetochore fibers at spindle poles throughout mitosis is prevented. Without centrosomes and NuMA, initial establishment of spindle microtubule focusing completely fails. Thus, NuMA is a defining feature of the mammalian spindle pole and functions as an essential tether linking bulk microtubules of the spindle to centrosomes.

Abbreviations used in this paper: 4-OHT, 4-hydroxytamoxifen; CENP-E, centromere protein E; ERTM, estrogen receptor tamoxifen mutant; ES, embryonic stem; Flp, flipase; FRT, Flp recombinase target; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; MEF, mouse embryo fibroblast; NuMA, nuclear mitotic apparatus; qPCR, quantitative PCR; STLC, S-trityl-L-cysteine; VAchT, vesicular acetylcholine transporter.

© 2009 Silk et al.
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