CENP-E Function at Kinetochores Is Essential for Chromosome Alignment

doi:10.1083/jcb.139.6.1373

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© The Rockefeller University Press, 0021-9525/1997//1373 $5.00
The Journal of Cell Biology, Volume 139, Number 6, , 1997 1373-1382

Article

CENP-E Function at Kinetochores Is Essential for Chromosome Alignment

B.T. Schaar^*, G.K.T. Chan, P. Maddox, E.D. Salmon, and T.J. Yen

^* Cell and Molecular Biology Graduate Group, University of Pennsylvania, Philadelphia, Pennsylvania 19103; Department of Biology, University of North Carolina, Chapel Hill, North Carolina 27599; and Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111

CENP-E is a kinesin-like protein that binds to kinetochoresand may provide functions that are critical for normal chromosomemotility during mitosis. To directly test the in vivo functionof CENP-E, we microinjected affinity-purified antibodies toblock the assembly of CENP-E onto kinetochores and then examinedthe behavior of these chromosomes. Chromosomes lacking CENP-Eat their kinetochores consistently exhibited two types of defectsthat blocked their alignment at the spindle equator. Chromosomes positioned near a pole remained mono-oriented as they wereunable to establish bipolar microtubule connections with theopposite pole. Chromosomes within the spindle established bipolarconnections that supported oscillations and normal velocitiesof kinetochore movement between the poles, but these bipolarconnections were defective because they failed to align thechromosomes into a metaphase plate.

Overexpression of a mutant that lacked the amino-terminal 803amino acids of CENP-E was found to saturate limiting bindingsites on kinetochores and competitively blocked endogenousCENP-E from assembling onto kinetochores. Chromosomes saturated with the truncated CENP-E mutant were never found to be alignedbut accumulated at the poles or were strewn within the spindleas was the case when cells were microinjected with CENP-E antibodies.As the motor domain was contained within the portion of CENP-Ethat was deleted, the chromosomal defect is likely attributedto the loss of motor function.

The combined data show that CENP-E provides kinetochore functionsthat are essential for monopolar chromosomes to establish bipolarconnections and for chromosomes with connections to both spindlepoles to align at the spindle equator. Both of these eventsrely on activities that are provided by CENP-E's motor domain.

Abbreviations used in this paper: ACA, anti-centromere autoimmune serum; DIC, differential interference contrast; GFP, green fluorescent protein; GST, glutathione-S-transferase; MBP, maltose-binding protein; VE, video enhanced.

Address all correspondence to Tim J. Yen, Fox Chase Cancer Center,770 Burholme Avenue, Philadelphia, PA 19111. Tel.: (215) 728-2590.Fax: (215) 728-3616.

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Gordon, M. B., Howard, L., Compton, D. A. (2001). Chromosome Movement in Mitosis Requires Microtubule Anchorage at Spindle Poles. JCB 152: 425-434 [Abstract] [Full Text]
Scaerou, F., Starr, D. A., Piano, F., Papoulas, O., Karess, R. E., Goldberg, M. L. (2001). The ZW10 and Rough Deal checkpoint proteins function together in a large, evolutionarily conserved complex targeted to the kinetochore. J. Cell Sci. 114: 3103-3114 [Abstract] [Full Text]
Ginkel, L. M., Wordeman, L. (2000). Expression and Partial Characterization of Kinesin-related Proteins in Differentiating and Adult Skeletal Muscle. Mol. Biol. Cell 11: 4143-4158 [Abstract] [Full Text]
Yu, H.-G., Dawe, R. K. (2000). Functional Redundancy in the Maize Meiotic Kinetochore. JCB 151: 131-142 [Abstract] [Full Text]
Yucel, J. K., Marszalek, J. D., McIntosh, J. R., Goldstein, L. S.B., Cleveland, D. W., Philp, A. V. (2000). CENP-meta, an Essential Kinetochore Kinesin Required for the Maintenance of Metaphase Chromosome Alignment in Drosophila. JCB 150: 1-12 [Abstract] [Full Text]
Pluta, A., Earnshaw, W., Goldberg, I. (2000). Interphase-specific association of intrinsic centromere protein CENP-C with HDaxx, a death domain-binding protein implicated in Fas-mediated cell death. J. Cell Sci. 111: 2029-2041 [Abstract]
Qiu, L., Burgess, A., Fairlie, D. P., Leonard, H., Parsons, P. G., Gabrielli, B. G. (2000). Histone Deacetylase Inhibitors Trigger a G2 Checkpoint in Normal Cells That Is Defective in Tumor Cells. Mol. Biol. Cell 11: 2069-2083 [Abstract] [Full Text]
Hunter, A., Wordeman, L (2000). How motor proteins influence microtubule polymerization dynamics. J. Cell Sci. 113: 4379-4389 [Abstract]
Starr, D., Saffery, R, Li, Z, Simpson, A., Choo, K., Yen, T., Goldberg, M. (2000). HZwint-1, a novel human kinetochore component that interacts with HZW10. J. Cell Sci. 113: 1939-1950 [Abstract]
McNally, K., Bazirgan, O., McNally, F. (2000). Two domains of p80 katanin regulate microtubule severing and spindle pole targeting by p60 katanin. J. Cell Sci. 113: 1623-1633 [Abstract]
Matthies, H. J.G., Messina, L. G., Namba, R., Greer, K. J., Walker, M.Y., Hawley, R. S. (1999). Mutations in the {alpha}-Tubulin 67C Gene Specifically Impair Achiasmate Segregation in Drosophila melanogaster. JCB 147: 1137-1144 [Abstract] [Full Text]
Sugata, N., Munekata, E., Todokoro, K. (1999). Characterization of a Novel Kinetochore Protein, CENP-H. J. Biol. Chem. 274: 27343-27346 [Abstract] [Full Text]
Chan, G.K.T., Jablonski, S.A., Sudakin, V., Hittle, J.C., Yen, T.J. (1999). Human Bubr1 Is a Mitotic Checkpoint Kinase That Monitors Cenp-E Functions at Kinetochores and Binds the Cyclosome/APC. JCB 146: 941-954 [Abstract] [Full Text]
Zhu, X. (1999). Structural Requirements and Dynamics of Mitosin-Kinetochore Interaction in M Phase. Mol. Cell. Biol. 19: 1016-1024 [Abstract] [Full Text]
Savoian, M. S., Earnshaw, W. C., Khodjakov, A., Rieder, C. L. (1999). Cleavage Furrows Formed between Centrosomes Lacking an Intervening Spindle and Chromosomes Contain Microtubule Bundles, INCENP, and CHO1 but Not CENP-E. Mol. Biol. Cell 10: 297-311 [Abstract] [Full Text]
Schumacher, J. M., Golden, A., Donovan, P. J. (1998). AIR-2: An Aurora/Ipl1-related Protein Kinase Associated with Chromosomes and Midbody Microtubules Is Required for Polar Body Extrusion and Cytokinesis in Caenorhabditis elegans Embryos. JCB 143: 1635-1646 [Abstract] [Full Text]
Chan, G.K.T., Schaar, B.T., Yen, T.J. (1998). Characterization of the Kinetochore Binding Domain of CENP-E Reveals Interactions with the Kinetochore Proteins CENP-F and hBUBR1. JCB 143: 49-63 [Abstract] [Full Text]
Zecevic, M., Catling, A. D., Eblen, S. T., Renzi, L., Hittle, J. C., Yen, T. J., Gorbsky, G. J., Weber, M. J. (1998). Active MAP Kinase in Mitosis: Localization at Kinetochores and Association with the Motor Protein CENP-E. JCB 142: 1547-1558 [Abstract] [Full Text]
Maney, T., Hunter, A. W., Wagenbach, M., Wordeman, L. (1998). Mitotic Centromere-associated Kinesin Is Important for Anaphase Chromosome Segregation. JCB 142: 787-801 [Abstract] [Full Text]
Mackay, A. M., Ainsztein, A. M., Eckley, D. M., Earnshaw, W. C. (1998). A Dominant Mutant of Inner Centromere Protein (INCENP), a Chromosomal Protein, Disrupts Prometaphase Congression and Cytokinesis. JCB 140: 991-1002 [Abstract] [Full Text]
Ashar, H. R., James, L., Gray, K., Carr, D., Black, S., Armstrong, L., Bishop, W. R., Kirschmeier, P. (2000). Farnesyl Transferase Inhibitors Block the Farnesylation of CENP-E and CENP-F and Alter the Association of CENP-E with the Microtubules. J. Biol. Chem. 275: 30451-30457 [Abstract] [Full Text]
Crespo, N. C., Ohkanda, J., Yen, T. J., Hamilton, A. D., Sebti, S. M. (2001). The Farnesyltransferase Inhibitor, FTI-2153, Blocks Bipolar Spindle Formation and Chromosome Alignment and Causes Prometaphase Accumulation during Mitosis of Human Lung Cancer Cells. J. Biol. Chem. 276: 16161-16167 [Abstract] [Full Text]
DeLuca, J. G., Newton, C. N., Himes, R. H., Jordan, M. A., Wilson, L. (2001). Purification and Characterization of Native Conventional Kinesin, HSET, and CENP-E from Mitotic HeLa Cells. J. Biol. Chem. 276: 28014-28021 [Abstract] [Full Text]
Howell, B.J., McEwen, B.F., Canman, J.C., Hoffman, D.B., Farrar, E.M., Rieder, C.L., Salmon, E.D. (2001). Cytoplasmic dynein/dynactin drives kinetochore protein transport to the spindle poles and has a role in mitotic spindle checkpoint inactivation. JCB 155: 1159-1172 [Abstract] [Full Text]