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Human CENP-I specifies localization of CENP-F, MAD1 and MAD2 to kinetochores and is essential for mitosis

Nature Cell Biology volume 5, pages 341–345 (2003)Cite this article

Abstract

The kinetochore, a macromolecular complex located at the centromere of chromosomes, provides essential functions for accurate chromosome segregation1,2. Kinetochores contain checkpoint proteins that monitor attachments between the kinetochore and microtubules to ensure that cells do not exit mitosis in the presence of unaligned chromosomes3,4. Here we report that human CENP-I, a constitutive protein of the kinetochore that shares limited similarity with Mis6 of Schizosaccharomyces pombe, is required for the localization of CENP-F and the checkpoint proteins MAD1 and MAD2 to kinetochores. Depletion of CENP-I from kinetochores causes the cell cycle to delay in G2. Although monopolar chromosomes in CENP-I-depleted cells fail to establish bipolar connections, the cells are unable to arrest in mitosis. These cells are transiently delayed in mitosis in a MAD2-dependent manner, even though their kinetochores are depleted of MAD2. The delay is extended considerably when the number of unattached kinetochores is increased. This suggests that no single unattached kinetochore in CENP-I-depleted cells can arrest mitosis. The collective output from many unattached kinetochores is required to reach a threshold signal of 'wait for anaphase' to sustain a prolonged mitotic arrest.

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Figure 1: Disruption of CENP-I delays progression through G2 and M.
Figure 2: CENP-I-depleted cells have monopolar chromosomes but do not arrest in mitosis.
Figure 3: Kinetochores depleted of CENP-I lack CENP-F, MAD1 and MAD2.
Figure 4: Nocodazole arrests CENP-I-depleted cells in mitosis.

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Acknowledgements

We thank G. Chan for reagents and advice, B. Conner for technical support and the core facilities at Fox Chase Cancer Center. S.T.L. is supported by Lawrence Greenwald Fellowship. This work was supported by grants from the NIH (GM44762, core grant CA06927) and an Appropriation from the Commonwealth of Pennsylvania.

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Authors and Affiliations

  1. Fox Chase Cancer Center, 7701 Burholme Avenue, Philadelphia, 19111, Pennsylvania, USA

    Song-Tao Liu, James C. Hittle, Sandra A. Jablonski, Michael S. Campbell & Tim J. Yen

  2. Bioscience Center, Nagoya University, Chikusa-ku, Nagoya, 464-8601, Japan

    Kinya Yoda

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Correspondence to Tim J. Yen.

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Supplementary information

Supplementary Figures

Figure S1 Human CENP-I is a constitutive kinetochore protein. (PDF 535 kb)

Figure S2 CENP-I antibody injection depleted CENP-I from kinetochores.

Figure S3 The kinetochore localization of BUB1, hZW10, hROD, CENP-E was not affected in CENP-I defective cells.

Figure S4 The protein levels of MAD1, MAD2 and CENP-F is unchanged in CENP-I siRNA treated cells.

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Liu, ST., Hittle, J., Jablonski, S. et al. Human CENP-I specifies localization of CENP-F, MAD1 and MAD2 to kinetochores and is essential for mitosis. Nat Cell Biol 5, 341–345 (2003). https://doi.org/10.1038/ncb953

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