The centromere-kinetochore complex: a repeat subunit model

doi:10.1083/jcb.113.5.1091

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The centromere-kinetochore complex: a repeat subunit model

RP Zinkowski, J Meyne and BR Brinkley
Department of Cell Biology, University of Alabama, Birmingham 35294.

The three-dimensional structure of the kinetochore and the DNA/protein composition of the centromere-kinetochore region was investigated using two novel techniques, caffeine-induced detachment of unreplicated kinetochores and stretching of kinetochores by hypotonic and/or shear forces generated in a cytocentrifuge. Kinetochore detachment was confirmed by EM and immunostaining with CREST autoantibodies. Electron microscopic analyses of serial sections demonstrated that detached kinetochores represented fragments derived from whole kinetochores. This was especially evident for the seven large kinetochores in the male Indian muntjac that gave rise to 80-100 fragments upon detachment. The kinetochore fragments, all of which interacted with spindle microtubules and progressed through the entire repertoire of mitotic movements, provide evidence for a subunit organization within the kinetochore. Further support for a repeat subunit model was obtained by stretching or uncoiling the metaphase centromere-kinetochore complex by hypotonic treatments. When immunostained with CREST autoantibodies and subsequently processed for in situ hybridization using synthetic centromere probes, stretched kinetochores displayed a linear array of fluorescent subunits arranged in a repetitive pattern along a centromeric DNA fiber. In addition to CREST antigens, each repetitive subunit was found to bind tubulin and contain cytoplasmic dynein, a microtubule motor localized in the zone of the corona. Collectively, the data suggest that the kinetochore, a plate-like structure seen by EM on many eukaryotic chromosomes is formed by the folding of a linear DNA fiber consisting of tandemly repeated subunits interspersed by DNA linkers. This model, unlike any previously proposed, can account for the structural and evolutional diversity of the kinetochore and its relationship to the centromere of eukaryotic chromosomes of many species.
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Marshall, O. J., Marshall, A. T., Choo, K.H. A. (2008). Three-dimensional localization of CENP-A suggests a complex higher order structure of centromeric chromatin. JCB 183: 1193-1202 [Abstract] [Full Text]
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Black, B. E., Brock, M. A., Bedard, S., Woods, V. L. Jr., Cleveland, D. W. (2007). An epigenetic mark generated by the incorporation of CENP-A into centromeric nucleosomes. Proc. Natl. Acad. Sci. USA 104: 5008-5013 [Abstract] [Full Text]
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Powell, K. (2006). The kinetochore uncoiled. JCB 173: 7-7 [Full Text]
Nakashima, H., Nakano, M., Ohnishi, R., Hiraoka, Y., Kaneda, Y., Sugino, A., Masumoto, H. (2005). Assembly of additional heterochromatin distinct from centromere-kinetochore chromatin is required for de novo formation of human artificial chromosome. J. Cell Sci. 118: 5885-5898 [Abstract] [Full Text]
Emanuele, M. J., McCleland, M. L., Satinover, D. L., Stukenberg, P. T. (2005). Measuring the Stoichiometry and Physical Interactions between Components Elucidates the Architecture of the Vertebrate Kinetochore. Mol. Biol. Cell 16: 4882-4892 [Abstract] [Full Text]
Nagaki, K., Kashihara, K., Murata, M. (2005). Visualization of Diffuse Centromeres with Centromere-Specific Histone H3 in the Holocentric Plant Luzula nivea. Plant Cell 17: 1886-1893 [Abstract] [Full Text]
Salmon, E.D, Cimini, D, Cameron, L.A, DeLuca, J.G (2005). Merotelic kinetochores in mammalian tissue cells. Phil Trans R Soc B 360: 553-568 [Abstract] [Full Text]
Pidoux, A. L, Allshire, R. C (2005). The role of heterochromatin in centromere function. Phil Trans R Soc B 360: 569-579 [Abstract] [Full Text]
Chueh, A. C., Wong, L. H., Wong, N., Choo, K.H. A. (2005). Variable and hierarchical size distribution of L1-retroelement-enriched CENP-A clusters within a functional human neocentromere. Hum Mol Genet 14: 85-93 [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]
Nakano, M., Okamoto, Y., Ohzeki, J.-i., Masumoto, H. (2003). Epigenetic assembly of centromeric chromatin at ectopic {alpha}-satellite sites on human chromosomes. J. Cell Sci. 116: 4021-4034 [Abstract] [Full Text]
Stear, J. H., Roth, M. B. (2002). Characterization of HCP-6, a C. elegans protein required to prevent chromosome twisting and merotelic attachment. Genes Dev. 16: 1498-1508 [Abstract] [Full Text]
Talbert, P. B., Masuelli, R., Tyagi, A. P., Comai, L., Henikoff, S. (2002). Centromeric Localization and Adaptive Evolution of an Arabidopsis Histone H3 Variant. Plant Cell 14: 1053-1066 [Abstract] [Full Text]
Ando, S., Yang, H., Nozaki, N., Okazaki, T., Yoda, K. (2002). CENP-A, -B, and -C Chromatin Complex That Contains the I-Type {alpha}-Satellite Array Constitutes the Prekinetochore in HeLa Cells. Mol. Cell. Biol. 22: 2229-2241 [Abstract] [Full Text]
Hagstrom, K. A., Holmes, V. F., Cozzarelli, N. R., Meyer, B. J. (2002). C. elegans condensin promotes mitotic chromosome architecture, centromere organization, and sister chromatid segregation during mitosis and meiosis. Genes Dev. 16: 729-742 [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]
Maggert, K. A., Karpen, G. H. (2001). The Activation of a Neocentromere in Drosophila Requires Proximity to an Endogenous Centromere. Genetics 158: 1615-1628 [Abstract] [Full Text]
Dernburg, A. F. (2001). Here, There, and Everywhere: Kinetochore Function on Holocentric Chromosomes. JCB 153: f33-f38 [Full Text]
Moore, L. L., Roth, M. B. (2001). Hcp-4, a Cenp-C-Like Protein inCaenorhabditis elegans, Is Required for Resolution of Sister Centromeres. JCB 153: 1199-1208 [Abstract] [Full Text]
Cimini, D., Howell, B., Maddox, P., Khodjakov, A., Degrassi, F., Salmon, E.D. (2001). Merotelic Kinetochore Orientation Is a Major Mechanism of Aneuploidy in Mitotic Mammalian Tissue Cells. JCB 153: 517-528 [Abstract] [Full Text]
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Yu, H.-G., Dawe, R. K. (2000). Functional Redundancy in the Maize Meiotic Kinetochore. JCB 151: 131-142 [Abstract] [Full Text]
Henikoff, S., Ahmad, K., Platero, J. S., van Steensel, B. (2000). From the Cover: Heterochromatic deposition of centromeric histone H3-like proteins. Proc. Natl. Acad. Sci. USA 97: 716-721 [Abstract] [Full Text]
Moore, L. L., Morrison, M., Roth, M. B. (1999). Hcp-1, a Protein Involved in Chromosome Segregation, Is Localized to the Centromere of Mitotic Chromosomes in Caenorhabditis elegans. JCB 147: 471-480 [Abstract] [Full Text]
Kaszás, E., Birchler, J. A. (1998). Meiotic Transmission Rates Correlate With Physical Features of Rearranged Centromeres in Maize. Genetics 150: 1683-1692 [Abstract] [Full Text]
Ananiev, E. V., Phillips, R. L., Rines, H. W. (1998). Chromosome-specific molecular organization of maize (Zea mays L.) centromeric regions. Proc. Natl. Acad. Sci. USA 95: 13073-13078 [Abstract] [Full Text]
Cambareri, E. B., Aisner, R., Carbon, J. (1998). Structure of the Chromosome VII Centromere Region in Neurospora crassa: Degenerate Transposons and Simple Repeats. Mol. Cell. Biol. 18: 5465-5477 [Abstract] [Full Text]
Vega, J. M., Feldman, M. (1998). Effect of the Pairing Gene Ph1 on Centromere Misdivision in Common Wheat. Genetics 148: 1285-1294 [Abstract] [Full Text]
Van Hooser, A, Goodrich, D., Allis, C., Brinkley, B., Mancini, M. (1998). Histone H3 phosphorylation is required for the initiation, but not maintenance, of mammalian chromosome condensation. J. Cell Sci. 111: 3497-3506 [Abstract]
Meluh, P. B., Koshland, D. (1997). Budding yeast centromere composition and assembly as revealed by in vivo�cross-linking. Genes Dev. 11: 3401-3412 [Abstract] [Full Text]
Ye, X., Sloboda, R. D. (1997). Molecular Characterization of p62, a Mitotic Apparatus Protein Required for Mitotic Progression. J. Biol. Chem. 272: 3606-3614 [Abstract] [Full Text]
Khodjakov, A., Cole, R. W., McEwen, B. F., Buttle, K. F., Rieder, C. L. (1997). Chromosome Fragments Possessing Only One Kinetochore Can Congress to the Spindle Equator. JCB 136: 229-240 [Abstract] [Full Text]
He, D, Brinkley, B. (1996). Structure and dynamic organization of centromeres/prekinetochores in the nucleus of mammalian cells. J. Cell Sci. 109: 2693-2704 [Abstract]
Brown, K., Wood, K., Cleveland, D. (1996). The kinesin-like protein CENP-E is kinetochore-associated throughout poleward chromosome segregation during anaphase-A. J. Cell Sci. 109: 961-969 [Abstract]
Zhu, X., Chang, K.-H., He, D., Mancini, M. A., Brinkley, W. R., Lee, W.-H. (1995). The C Terminus of Mitosin Is Essential for Its Nuclear Localization, Centromere/Kinetochore Targeting, and Dimerization. J. Biol. Chem. 270: 19545-19550 [Abstract] [Full Text]
Mitchell, A., Nicol, L, Malloy, P, Kipling, D (1993). Novel structural organisation of a Mus musculus DBA/2 chromosome shows a fixed position for the centromere. J. Cell Sci. 106: 79-85 [Abstract]
Ouspenski, I., Brinkley, B. (1993). Centromeric DNA cloned from functional kinetochore fragments in mitotic cells with unreplicated genomes. J. Cell Sci. 105: 359-367 [Abstract]
Nicklas, R., Krawitz, L., Ward, S. (1993). Odd chromosome movement and inaccurate chromosome distribution in mitosis and meiosis after treatment with protein kinase inhibitors. J. Cell Sci. 104: 961-973 [Abstract]
Clarke, L., Baum, M., Marschall, L.G., Ngan, V.K., Steiner, N.C. (1993). Structure and Function of Schizosaccharomyces pombe Centromeres. Cold Spring Harb Symp Quant Biol 58: 687-695 [Abstract]
(1992). Abstracts. Toxicol Pathol 20: 655-670
Earnshaw, W.C., Bernat, R.L., Cooke, C.A., Rothfield, N.F. (1991). Role of the Centromere/Kinetochore in Cell Cycle Control. Cold Spring Harb Symp Quant Biol 56: 675-685 [Abstract]