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A more recent version of this article appeared on January 1, 2005.
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Submitted on October 11, 2004
Revised on November 23, 2004
Accepted on November 23, 2004

Proteome analysis of the human mitotic spindle

G. Sauer, R. Koerner, A. Hanisch, A. Ries, E. A. Nigg, and H. H.W. Sillje

Cell Biology, Max Planck Institute for Biochemistry, Martinsried, Bayern D-82152

Corresponding Author: sillje{at}biochem.mpg.de

The accurate distribution of sister chromatids during cell division is crucial for the generation of two cells with the same complement of genetic information. A highly dynamic microtubule based structure, the mitotic spindle, carries out the physical separation of the chromosomes to opposite poles of the cells and, moreover, determines the cell division cleavage plane. In animal cells the spindle comprises microtubules that radiate from the microtubule organizing centers, the centrosomes, and interact with kinetochores on the chromosomes. Malfunctioning of the spindle can lead to chromosome missegregation and hence result in aneuploidy, a hallmark of most human cancers. Despite major progress in deciphering the temporal and spatial regulation of the mitotic spindle, its composition and function are not fully understood. A more complete inventory of spindle components would therefore constitute an important advance. Here we describe the purification of human mitotic spindles and their analysis by tandem mass spectrometry. We identified 151 proteins previously known to associate with the spindle apparatus, centrosomes and/or kinetochores and 644 other proteins, including 154 uncharacterized components that did not show obvious homologies to known proteins and did not contain motifs indicative of a particular localization. Of these uncharacterized proteins, 17 were tagged and localized in transfected mitotic cells, resulting in the identification of 6 genuine spindle components (KIAA0008, CdcA8, KIAA1187, FLJ12649, FLJ90806 and C20Orf129). This study illustrates the strength of a proteomic approach for the analysis of isolated human spindles and identifies several novel spindle components for future functional studies.


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