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Published online 22 January 2001. doi:10.1083/jcb.152.2.349
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© The Rockefeller University Press, 0021-9525/2001//349 $5.00
The Journal of Cell Biology, Volume 152, Number 2, , 2001 349-360

Original Article

The Ndc80p Complex from Saccharomyces cerevisiae Contains Conserved Centromere Components and Has a Function in Chromosome Segregation



Philip A. Wiggea and John V. Kilmartina

a Medical Research Council, Laboratory of Molecular Biology, Cambridge CB2 2QH, United Kingdom
MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK.44-1223-412-14244-1223-402-242

jvk{at}mrc-lmb.cam.ac.uk

We have purified a complex from Saccharomyces cerevisiae containing the spindle components Ndc80p, Nuf2p, Spc25p, and Spc24p. Temperature-sensitive mutants in NDC80, SPC25, and SPC24 show defects in chromosome segregation. In spc24-1 cells, green fluorescence protein (GFP)-labeled centromeres fail to split during spindle elongation, and in addition some centromeres may detach from the spindle. Chromatin immunoprecipitation assays show an association of all four components of the complex with the yeast centromere. Homologues of Ndc80p, Nuf2p, and Spc24p were found in Schizosaccharomyces pombe and GFP tagging showed they were located at the centromere. A human homologue of Nuf2p was identified in the expressed sequence tag database. Immunofluorescent staining with anti–human Nuf2p and with anti-HEC, the human homologue of Ndc80p, showed that both proteins are at the centromeres of mitotic HeLa cells. Thus the Ndc80p complex contains centromere-associated components conserved between yeasts and vertebrates.

Key Words: Ndc80p • Nuf2p • Spc25p • Spc24p • centromere

© 2001 The Rockefeller University Press

Philip A. Wigge's present address is Plant Biology Laboratory, The Salk Institute for Biological Studies, La Jolla, CA 92037.

Abbreviations used in this paper: CBF, centromere-binding factor; ChIP, chromatin immunoprecipitation; GFP, green fluorescent protein; GST, glutathione S-transferase; hNuf2R, human Nuf2p-related; MALDI, matrix-assisted laser desorption/ionization; prA, protein A; SPB, spindle pole body; ts, temperature-sensitive; wt, wild-type.


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