J. Cell Biol.
© The Rockefeller University Press
0021-9525/97/12/1373/10 $2.00
Volume 139, Number 6, December 15, 1997 1373-1382

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 kinetochores and may provide functions that are critical for normal chromosome motility during mitosis. To directly test the in vivo function of CENP-E, we microinjected affinity-purified antibodies to block the assembly of CENP-E onto kinetochores and then examined the behavior of these chromosomes. Chromosomes lacking CENP-E at their kinetochores consistently exhibited two types of defects that blocked their alignment at the spindle equator. Chromosomes positioned near a pole remained mono-oriented as they were unable to establish bipolar microtubule connections with the opposite pole. Chromosomes within the spindle established bipolar connections that supported oscillations and normal velocities of kinetochore movement between the poles, but these bipolar connections were defective because they failed to align the chromosomes into a metaphase plate.

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

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

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

• Kong, X., Ball, A. R. Jr., Sonoda, E., Feng, J., Takeda, S., Fukagawa, T., Yen, T. J., Yokomori, K. (2009). Cohesin Associates with Spindle Poles in a Mitosis-specific Manner and Functions in Spindle Assembly in Vertebrate Cells. Mol. Biol. Cell 20: 1289-1301 [Abstract] [Full Text]  
• Watanabe, N., Sekine, T., Takagi, M., Iwasaki, J.-i., Imamoto, N., Kawasaki, H., Osada, H. (2009). Deficiency in Chromosome Congression by the Inhibition of Plk1 Polo Box Domain-dependent Recognition. J. Biol. Chem. 284: 2344-2353 [Abstract] [Full Text]  
• Henderson, M. C., Shaw, Y.-J. Y., Wang, H., Han, H., Hurley, L. H., Flynn, G., Dorr, R. T., Von Hoff, D. D. (2009). UA62784, a novel inhibitor of centromere protein E kinesin-like protein. Molecular Cancer Therapeutics 8: 36-44 [Abstract] [Full Text]  
• Wood, K. W., Chua, P., Sutton, D., Jackson, J. R. (2008). Centromere-Associated Protein E: A Motor That Puts the Brakes on the Mitotic Checkpoint. Clin. Cancer Res. 14: 7588-7592 [Abstract] [Full Text]  
• Huang, H., Hittle, J., Zappacosta, F., Annan, R. S., Hershko, A., Yen, T. J. (2008). Phosphorylation sites in BubR1 that regulate kinetochore attachment, tension, and mitotic exit. JCB 183: 667-680 [Abstract] [Full Text]  
• Kim, Y., Heuser, J. E., Waterman, C. M., Cleveland, D. W. (2008). CENP-E combines a slow, processive motor and a flexible coiled coil to produce an essential motile kinetochore tether. JCB 181: 411-419 [Abstract] [Full Text]  
• Yardimci, H., van Duffelen, M., Mao, Y., Rosenfeld, S. S., Selvin, P. R. (2008). The mitotic kinesin CENP-E is a processive transport motor. Proc. Natl. Acad. Sci. USA 105: 6016-6021 [Abstract] [Full Text]  
• Dalton, W. B., Nandan, M. O., Moore, R. T., Yang, V. W. (2007). Human Cancer Cells Commonly Acquire DNA Damage during Mitotic Arrest. Cancer Res. 67: 11487-11492 [Abstract] [Full Text]  
• Liu, J., Desai, A., Onuchic, J. N., Hwa, T. (2007). A mechanobiochemical mechanism for monooriented chromosome oscillation in mitosis. Proc. Natl. Acad. Sci. USA 104: 16104-16109 [Abstract] [Full Text]  
• Liu, D., Ding, X., Du, J., Cai, X., Huang, Y., Ward, T., Shaw, A., Yang, Y., Hu, R., Jin, C., Yao, X. (2007). Human NUF2 Interacts with Centromere-associated Protein E and Is Essential for a Stable Spindle Microtubule-Kinetochore Attachment. J. Biol. Chem. 282: 21415-21424 [Abstract] [Full Text]  
• Grishchuk, E. L., Spiridonov, I. S., McIntosh, J. R. (2007). Mitotic Chromosome Biorientation in Fission Yeast Is Enhanced by Dynein and a Minus-end-directed, Kinesin-like Protein. Mol. Biol. Cell 18: 2216-2225 [Abstract] [Full Text]  
• Schafer-Hales, K., Iaconelli, J., Snyder, J. P., Prussia, A., Nettles, J. H., El-Naggar, A., Khuri, F. R., Giannakakou, P., Marcus, A. I. (2007). Farnesyl transferase inhibitors impair chromosomal maintenance in cell lines and human tumors by compromising CENP-E and CENP-F function. Molecular Cancer Therapeutics 6: 1317-1328 [Abstract] [Full Text]  
• Kanta, H., Laprade, L., Almutairi, A., Pinto, I. (2006). Suppressor Analysis of a Histone Defect Identifies a New Function for the Hda1 Complex in Chromosome Segregation. Genetics 173: 435-450 [Abstract] [Full Text]  
• Kapoor, T. M., Lampson, M. A., Hergert, P., Cameron, L., Cimini, D., Salmon, E. D., McEwen, B. F., Khodjakov, A. (2006). Chromosomes Can Congress to the Metaphase Plate Before Biorientation. Science 311: 388-391 [Abstract] [Full Text]  
• Hannak, E., Heald, R. (2006). Xorbit/CLASP links dynamic microtubules to chromosomes in the Xenopus meiotic spindle. JCB 172: 19-25 [Abstract] [Full Text]  
• Mao, Y., Desai, A., Cleveland, D. W. (2005). Microtubule capture by CENP-E silences BubR1-dependent mitotic checkpoint signaling. JCB 170: 873-880 [Abstract] [Full Text]  
• Zhu, C., Zhao, J., Bibikova, M., Leverson, J. D., Bossy-Wetzel, E., Fan, J.-B., Abraham, R. T., Jiang, W. (2005). Functional Analysis of Human Microtubule-based Motor Proteins, the Kinesins and Dyneins, in Mitosis/Cytokinesis Using RNA Interference. Mol. Biol. Cell 16: 3187-3199 [Abstract] [Full Text]  
• Minderman, H., Conroy, J. M., O'Loughlin, K. L., McQuaid, D., Quinn, P., Li, S., Pendyala, L., Nowak, N. J., Baer, M. R. (2005). In vitro and in vivo irinotecan-induced changes in expression profiles of cell cycle and apoptosis-associated genes in acute myeloid leukemia cells. Molecular Cancer Therapeutics 4: 885-900 [Abstract] [Full Text]  
• Dai, J., Sultan, S., Taylor, S. S., Higgins, J. M.G. (2005). The kinase haspin is required for mitotic histone H3 Thr 3 phosphorylation and normal metaphase chromosome alignment. Genes Dev. 19: 472-488 [Abstract] [Full Text]  
• Maiato, H., DeLuca, J., Salmon, E. D., Earnshaw, W. C. (2004). The dynamic kinetochore-microtubule interface. J. Cell Sci. 117: 5461-5477 [Abstract] [Full Text]  
• Stear, J. H., Roth, M. B. (2004). The Caenorhabditis elegans Kinetochore Reorganizes at Prometaphase and in Response to Checkpoint Stimuli. Mol. Biol. Cell 15: 5187-5196 [Abstract] [Full Text]  
• Tanudji, M., Shoemaker, J., L'Italien, L., Russell, L., Chin, G., Schebye, X. M. (2004). Gene Silencing of CENP-E by Small Interfering RNA in HeLa Cells Leads to Missegregation of Chromosomes after a Mitotic Delay. Mol. Biol. Cell 15: 3771-3781 [Abstract] [Full Text]  
• Levesque, A. A., Howard, L., Gordon, M. B., Compton, D. A. (2003). A Functional Relationship between NuMA and Kid Is Involved in Both Spindle Organization and Chromosome Alignment in Vertebrate Cells. Mol. Biol. Cell 14: 3541-3552 [Abstract] [Full Text]  
• Weaver, B. A.A., Bonday, Z. Q., Putkey, F. R., Kops, G. J.P.L., Silk, A. D., Cleveland, D. W. (2003). Centromere-associated protein-E is essential for the mammalian mitotic checkpoint to prevent aneuploidy due to single chromosome loss. JCB 162: 551-563 [Abstract] [Full Text]  
• Liu, S.-T., Chan, G. K.T., Hittle, J. C., Fujii, G., Lees, E., Yen, T. J. (2003). Human MPS1 Kinase Is Required for Mitotic Arrest Induced by the Loss of CENP-E from Kinetochores. Mol. Biol. Cell 14: 1638-1651 [Abstract] [Full Text]  
• Harper, J. W., Burton, J. L., Solomon, M. J. (2002). The anaphase-promoting complex: it's not just for mitosis any more. Genes Dev. 16: 2179-2206 [Full Text]  
• Kapoor, T. M., Compton, D. A. (2002). Searching for the middle ground: mechanisms of chromosome alignment during mitosis. JCB 157: 551-556 [Abstract] [Full Text]  
• Zhou, J., Panda, D., Landen, J. W., Wilson, L., Joshi, H. C. (2002). Minor Alteration of Microtubule Dynamics Causes Loss of Tension across Kinetochore Pairs and Activates the Spindle Checkpoint. J. Biol. Chem. 277: 17200-17208 [Abstract] [Full Text]  
• Bloom, K. (2002). Yeast weighs in on the elusive spindle matrix: New filaments in the nucleus. Proc. Natl. Acad. Sci. USA 99: 4757-4759 [Full Text]  
• Taylor, S. S., Hussein, D., Wang, Y., Elderkin, S., Morrow, C. J. (2002). Kinetochore localisation and phosphorylation of the mitotic checkpoint components Bub1 and BubR1 are differentially regulated by spindle events in human cells. J. Cell Sci. 114: 4385-4395 [Abstract] [Full Text]  
• Eaker, S., Pyle, A., Cobb, J., Handel, M. A. (2002). Evidence for meiotic spindle checkpoint from analysis of spermatocytes from Robertsonian-chromosome heterozygous mice. J. Cell Sci. 114: 2953-2965 [Abstract] [Full Text]  
• Hussein, D., Taylor, S. S. (2002). Farnesylation of Cenp-F is required for G2/M progression and degradation after mitosis. J. Cell Sci. 115: 3403-3414 [Abstract] [Full Text]  
• West, R. R., Malmstrom, T., McIntosh, J. R. (2002). Kinesins klp5+ and klp6+ are required for normal chromosome movement in mitosis. J. Cell Sci. 115: 931-940 [Abstract] [Full Text]  
• Jullien, D., Vagnarelli, P., Earnshaw, W. C., Adachi, Y. (2002). Kinetochore localisation of the DNA damage response component 53BP1 during mitosis. J. Cell Sci. 115: 71-79 [Abstract] [Full Text]  
• Levesque, A. A., Compton, D. A. (2001). The chromokinesin Kid is necessary for chromosome arm orientation and oscillation, but not congression, on mitotic spindles. JCB 154: 1135-1146 [Abstract] [Full Text]  
• McEwen, B. F., Chan, G. K.T., Zubrowski, B., Savoian, M. S., Sauer, M. T., Yen, T. J. (2001). CENP-E Is Essential for Reliable Bioriented Spindle Attachment, but Chromosome Alignment Can Be Achieved via Redundant Mechanisms in Mammalian Cells. Mol. Biol. Cell 12: 2776-2789 [Abstract] [Full Text]  
• Topper, L. M., Bastians, H., Ruderman, J. V., Gorbsky, G. J. (2001). Elevating the level of Cdc34/Ubc3 ubiquitin-conjugating enzyme in mitosis inhibits association of CENP-E with kinetochores and blocks the metaphase alignment of chromosomes. JCB 154: 707-718 [Abstract] [Full Text]  
• Hoffman, D. B., Pearson, C. G., Yen, T. J., Howell, B. J., Salmon, E.D. (2001). Microtubule-dependent Changes in Assembly of Microtubule Motor Proteins and Mitotic Spindle Checkpoint Proteins at PtK1 Kinetochores. Mol. Biol. Cell 12: 1995-2009 [Abstract] [Full Text]  
• Sharp-Baker, H., Chen, R.-H. (2001). Spindle Checkpoint Protein Bub1 Is Required for Kinetochore Localization of Mad1, Mad2, Bub3, and CENP-E, Independently of Its Kinase Activity. JCB 153: 1239-1250 [Abstract] [Full Text]  
• Fontijn, R. D., Goud, B., Echard, A., Jollivet, F., van Marle, J., Pannekoek, H., Horrevoets, A. J. G. (2001). The Human Kinesin-Like Protein RB6K Is under Tight Cell Cycle Control and Is Essential for Cytokinesis. Mol. Cell. Biol. 21: 2944-2955 [Abstract] [Full Text]  
• 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]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents