Abstract
The kinetochore is a complex molecular machine that directs chromosome segregation during mitosis. It is one of the most elaborate subcellular protein structures in eukaryotes, comprising more than 100 different proteins. Inner kinetochore proteins associate with specialized centromeric chromatin containing the histone H3 variant centromere protein A (CENP-A) in place of H3. Outer kinetochore proteins bind to microtubules and signal to delay anaphase onset when microtubules are absent. Since the first kinetochore proteins were discovered and cloned 30 years ago using autoimmune sera from patients with scleroderma-spectrum disease, much has been learnt about the composition, functions and regulation of this remarkable structure.
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References
Fukagawa, T. & Earnshaw, W. C. The centromere: chromatin foundation for the kinetochore machinery. Dev. Cell 30, 496–508 (2014).
Shang, W. H. et al. Chickens possess centromeres with both extended tandem repeats and short non- tandem-repetitive sequences. Genome Res. 20, 1219–1228 (2010).
Earnshaw, W. C. et al. Esperanto for histones: CENP-A, not CenH3, is the centromeric histone H3 variant. Chromosome Res. 21, 101–106 (2013).
De Rop, V., Padeganeh, A. & Maddox, P. S. CENP-A: the key player behind centromere identity, propagation, and kinetochore assembly. Chromosoma 121, 527–538 (2012).
Skene, P. J. & Henikoff, S. Histone variants in pluripotency and disease. Development 140, 2513–2524 (2013).
Bloom, K. S. Centromeric heterochromatin: the primordial segregation machine. Annu. Rev. Genet. 48, 457–484 (2014).
Blower, M. D., Sullivan, B. A. & Karpen, G. H. Conserved organization of centromeric chromatin in flies and humans. Dev. Cell 2, 319–330 (2002).
Carroll, C. W., Silva, M. C., Godek, K. M., Jansen, L. E. & Straight, A. F. Centromere assembly requires the direct recognition of CENP-A nucleosomes by CENP-N. Nature Cell Biol. 11, 896–902 (2009).
Carroll, C. W., Milks, K. J. & Straight, A. F. Dual recognition of CENP-A nucleosomes is required for centromere assembly. J. Cell Biol. 189, 1143–1155 (2010).
Fachinetti, D. et al. A two-step mechanism for epigenetic specification of centromere identity and function. Nature Cell Biol. 15, 1056–1066 (2013).
Hudson, D. et al. Centromere protein B null mice are mitotically and meiotically normal but have lower body and testis weights. J. Cell Biol. 141, 309–319 (1998).
Perez-Castro, A. V. et al. Centromeric protein B null mice are viable with no apparent abnormalities. Dev. Biol. 201, 135–143 (1998).
Kapoor, M. et al. The cenpB gene is not essential in mice. Chromosoma 107, 570–576 (1998).
Moroi, Y., Peebles, C., Fritzler, M. J., Steigerwald, J. & Tan, E. M. Autoantibody to centromere (kinetochore) in scleroderma sera. Proc. Natl Acad. Sci. USA 77, 1627–1631 (1980).
Moroi, Y., Hartman, A. L., Nakane, P. K. & Tan, E. M. Distribution of kinetochore (centromere) antigen in mammalian cell nuclei. J. Cell Biol. 90, 254–259 (1981).
Brenner, S., Pepper, D., Berns, M. W., Tan, E. & Brinkley, B. R. Kinetochore structure, duplication and distribution in mammalian cells: analysis by human autoantibodies from scleroderma patients. J. Cell Biol. 91, 95–102 (1981).
Brinkley, B. R., Valdivia, M. M., Tousson, A. & Brenner, S. L. Compound kinetochores of the Indian muntjac: evolution by linear fusion of unit kinetochores. Chromosoma 91, 1–11 (1984).
Earnshaw, W. C., Halligan, N., Cooke, C. & Rothfield, N. The kinetochore is part of the chromosome scaffold. J. Cell Biol. 98, 352–357 (1984).
Earnshaw, W. C. & Rothfield, N. Identification of a family of human centromere proteins using autoimmune sera from patients with scleroderma. Chromosoma 91, 313–321 (1985).
Guldner, H. H., Lakomek, H.-J. & Bautz, F. A. Human anti-centromere sera recognise a 19.5 kD non-histone chromosomal protein from HeLa cells. Clin. Exp. Immunol. 58, 13–20 (1984).
Young, R. A. & Davis, R. B. Yeast polymerase II genes: isolation with antibody probes. Science 222, 778–782 (1983).
Earnshaw, W. C. et al. Molecular cloning of cDNA for CENP-B, the major human centromere autoantigen. J. Cell Biol. 104, 817–829 (1987).
Saitoh, H. et al. CENP-C, an autoantigen in scleroderma, is a component of the human inner kinetochore plate. Cell 70, 115–125 (1992).
Palmer, D. K. & Margolis, R. L. A 17-kD centromere protein (CENP-A) copurifies with nucleosome core particles and with histones. J. Cell. Biol. 104, 805–815 (1987).
Sullivan, K. F., Hechenberger, M. & Masri, K. Human CENP-A contains a histone H3 related histone fold domain that is required for targeting to the centromere. J. Cell Biol. 127, 581–192 (1994).
Ando, S., Yang, H., Nozaki, N., Okazaki, T. & Yoda, K. CENP-A, -B, and -C chromatin complex that contains the I-type α-satellite array constitutes the prekinetochore in HeLa cells. Mol. Cell. Biol. 22, 2229–2241 (2002).
Foltz, D. R. et al. The human CENP-A centromeric nucleosome-associated complex. Nature Cell Biol. 8, 458–469 (2006).
Hori, T., Okada, M., Maenaka, K. & Fukagawa, T. CENP-O class proteins form a stable complex and are required for proper kinetochore function. Mol. Biol. Cell 19, 843–854 (2008).
Santaguida, S. & Musacchio, A. The life and miracles of kinetochores. EMBO J. 28, 2511–2531 (2009).
Perpelescu, M. & Fukagawa, T. The ABCs of CENPs. Chromosoma 120, 425–446 (2011).
Biggins, S. The composition, functions, and regulation of the budding yeast kinetochore. Genetics 194, 817–846 (2013).
Westhorpe, F. G. & Straight, A. F. Functions of the centromere and kinetochore in chromosome segregation. Curr. Opin. Cell Biol. 25, 334–340 (2013).
Cheerambathur, D. K. & Desai, A. Linked in: formation and regulation of microtubule attachments during chromosome segregation. Curr. Opin. Cell Biol. 26, 113–122 (2014).
Stoler, S., Keith, K. C., Curnick, K. E. & Fitzgerald-Hayes, M. A mutation in CSE4, an essential gene encoding a novel chromatin-associated protein in yeast, causes chromosome nondisjunction and cell cycle arrest at mitosis. Genes Dev. 9, 573–586 (1995).
Takahashi, K., Chen, E. S. & Yanagida, M. Requirement of Mis6 centromere connector for localizing a CENP-A-like protein in fission yeast. Science 288, 2215–2219 (2000).
Henikoff, S. & Furuyama, T. The unconventional structure of centromeric nucleosomes. Chromosoma 121, 341–352 (2012).
Kurumizaka, H., Horikoshi, N., Tachiwana, H. & Kagawa, W. Current progress on structural studies of nucleosomes containing histone H3 variants. Curr. Opin. Struct. Biol. 23, 109–115 (2012).
Padeganeh, A., De Rop, V. & Maddox, P. S. Nucleosomal composition at the centromere: a numbers game. Chromosome Res. 21, 27–36 (2013).
Catania, S. & Allshire, R. C. Anarchic centromeres: deciphering order from apparent chaos. Curr. Opin. Cell Biol. 26, 41–50 (2014).
Foltz, D. R. et al. Centromere-specific assembly of CENP-A nucleosomes is mediated by HJURP. Cell 137, 472–484 (2009).
Dunleavy, E. M. et al. HJURP is a cell-cycle- dependent maintenance and deposition factor of CENP-A at centromeres. Cell 137, 485–497 (2009).
Jansen, L. E., Black, B. E., Foltz, D. R. & Cleveland, D. W. Propagation of centromeric chromatin requires exit from mitosis. J. Cell Biol. 176, 795–805 (2007).
Howman, E. V. et al. Early disruption of centromeric chromatin organization in centromere protein A (Cenpa) null mice. Proc. Natl Acad. Sci. USA 97, 1148–1153 (2000).
Oegema, K., Desai, A., Rybina, S., Kirkham, M. & Hyman, A. A. Functional analysis of kinetochore assembly in Caenorhabditis elegans. J. Cell Biol. 153, 1209–1226 (2001).
Heun, P. et al. Mislocalization of the Drosophila centromere-specific histone CID promotes formation of functional ectopic kinetochores. Dev. Cell 10, 303–315 (2006).
Barnhart, M. C. et al. HJURP is a CENP-A chromatin assembly factor sufficient to form a functional de novo kinetochore. J. Cell Biol. 194, 229–243 (2011).
Tudor, M., Lobocka, M., Goodell, M., Pettitt, J. & O'Hare, K. The pogo transposable element family of Drosophila melanogaster. Mol. Gen. Genet. 232, 126–134 (1992).
Casola, C., Hucks, D. & Feschotte, C. Convergent domestication of pogo-like transposases into centromere-binding proteins in fission yeast and mammals. Mol. Biol. Evol. 25, 29–41 (2008).
Cooke, C. A., Bernat, R. L. & Earnshaw, W. C. CENP-B: a major human centromere protein located beneath the kinetochore. J. Cell Biol. 110, 1475–1488 (1990).
Masumoto, H., Masukata, H., Muro, Y., Nozaki, N. & Okazaki, T. A human centromere antigen (CENP-B) interacts with a short specific sequence in alphoid DNA, a human centromeric satellite. J. Cell Biol. 109, 1963–1973 (1989).
Okada, T. et al. CENP-B controls centromere formation depending on the chromatin context. Cell 131, 1287–1300 (2007).
Liu, S. T., Rattner, J. B., Jablonski, S. A. & Yen, T. J. Mapping the assembly pathways that specify formation of the trilaminar kinetochore plates in human cells. J. Cell Biol. 175, 41–53 (2006).
Erhardt, S. et al. Genome-wide analysis reveals a cell cycle-dependent mechanism controlling centromere propagation. J. Cell Biol. 183, 805–818 (2008).
Brown, M. T. Sequence similarities between the yeast chromosome segregation protein Mif2 and the human centromere protein CENP-C. Gene 160, 111–116 (1995).
Meluh, P. B. & Koshland, D. Evidence that the MIF2 gene of Saccharomyces cerevisiae encodes a centromere protein with homology to the mammalian centromere protein CENP-C. Mol. Biol. Cell 6, 793–807 (1995).
Sugimoto, K., Yata, H., Muro, Y. & Himeno, M. Human centromere protein C (CENP-C) is a DNA-binding protein which possesses a novel DNA-binding motif. J. Biochem. 116, 877–881 (1994).
Yang, C. H., Tomkiel, J., Saitoh, H., Johnson, D. H. & Earnshaw, W. C. Identification of overlapping DNA-binding and centromere-targeting domains in the human kinetochore protein CENP-C. Mol. Cell. Biol. 16, 3576–3586 (1996).
Kato, H. et al. A conserved mechanism for centromeric nucleosome recognition by centromere protein CENP-C. Science 340, 1110–1113 (2013).
Moree, B., Meyer, C. B., Fuller, C. J. & Straight, A. F. CENP-C recruits M18BP1 to centromeres to promote CENP-A chromatin assembly. J. Cell Biol. 194, 855–871 (2011).
Cheeseman, I. M., Chappie, J. S., Wilson-Kubalek, E. M. & Desai, A. The conserved KMN network constitutes the core microtubule-binding site of the kinetochore. Cell 127, 983–997 (2006).
Przewloka, M. R. et al. CENP-C is a structural platform for kinetochore assembly. Curr. Biol. 21, 399–405 (2011).
Screpanti, E. et al. Direct binding of Cenp-C to the Mis12 complex joins the inner and outer kinetochore. Curr. Biol. 21, 391–398 (2011).
Wigge, P. A. & Kilmartin, J. V. The Ndc80p complex from Saccharomyces cerevisiae contains conserved centromere components and has a function in chromosome segregation. J. Cell Biol. 152, 349–360 (2001).
He, X., Rines, D. R., Espelin, C. W. & Sorger, P. K. Molecular analysis of kinetochore-microtubule attachment in budding yeast. Cell 106, 195–206 (2001).
De Wulf, P., McAinsh, A. D. & Sorger, P. K. Hierarchical assembly of the budding yeast kinetochore from multiple subcomplexes. Genes Dev. 17, 2902–2921 (2003).
Cheeseman, I. M. et al. A conserved protein network controls assembly of the outer kinetochore and its ability to sustain tension. Genes Dev. 18, 2255–2268 (2004).
DeLuca, J. et al. Hec1 and nuf2 are core components of the kinetochore outer plate essential for organizing microtubule attachment sites. Mol. Biol. Cell. 16, 519–531 (2005).
Wiener, E. S. et al. Clinical associations of anticentromere antibodies and antibodies to topoisomerase I: a study of 355 patients. Arthritis Rheum. 31, 378–385 (1988).
Wiener, E. S. et al. Prognostic significance of anticentromere antibodies and topoisomerase I antibodies in Raynaud's disease: a prospective study. Arthritis Rheum. 34, 68–77 (1989).
Earnshaw, W. C., Machlin, P. S., Bordwell, B., Rothfield, N. F. & Cleveland, D. W. Analysis of anti-centromere autoantibodies using cloned autoantigen CENP-B. Proc. Natl Acad. Sci. USA 84, 4979–4983 (1987).
Warburton, P. E. et al. Immunolocalization of CENP-A suggests a distinct nucleosome structure at the inner kinetochore plate of active centromeres. Curr. Biol. 7, 901–904 (1997).
Metzner, R. Beiträge zur Granulalehre. I. Kern und kerntheilung. Arch. Anat. Physiol. 309–348 (in German) (1894).
Darlington, C. D. The external mechanics of the chromosomes. I — the scope of enquiry. Proc. R. Soc. Lond. B 121, 264–273 (1936).
Luykx, P. The structure of the kinetochore in meiosis and mitosis in Urechis eggs. Exp. Cell Res. 39, 643–657 (1965).
Brinkley, B. R. & Stubblefield, E. The fine structure of the kinetochore of a mammalian cell in vitro. Chromosoma 19, 28–43 (1966).
Jokelainen, P. T. The ultrastructure and spatial organization of the metaphase kinetochore in mitotic rat cells. J. Ultrastruct. Res. 19, 19–44 (1967).
Earnshaw, W. C. & Migeon, B. A family of centromere proteins is absent from the latent centromere of a stable isodicentric chromosome. Chromosoma 92, 290–296 (1985).
Voullaire, L. E., Slater, H. R., Petrovic, V. & Choo, K. H. A functional marker centromere with no detectible α-satellite, satellite III, or CENP-B protein: activation of a latent centromere? Am. J. Hum. Genet. 52, 1153–1163 (1993).
Sullivan, B. A. & Karpen, G. H. Centromeric chromatin exhibits a histone modification pattern that is distinct from both euchromatin and heterochromatin. Nature Struct. Mol. Biol. 11, 1076–1083 (2004).
Cheeseman, I. M. & Desai, A. Molecular architecture of the kinetochore-microtubule interface. Nature Rev. Mol. Cell. Biol. 9, 33–46 (2008).
Nakano, M. et al. Inactivation of a human kinetochore by specific targeting of chromatin modifiers. Dev. Cell 14, 507–522 (2008).
Tachiwana, H. et al. Crystal structure of the human centromeric nucleosome containing CENP-A. Nature 476, 232–235 (2011).
Hori, T., Shang, W. H., Takeuchi, K. & Fukagawa, T. The CCAN recruits CENP-A to the centromere and forms the structural core for kinetochore assembly. J. Cell Biol. 200, 45–60 (2013).
Acknowledgements
The studies described here are testimony to the power of scientific collaborations. This story would not have developed without N. Rothfield, D. Cleveland, K. Sullivan and T. Pollard, who provided inspiration, reagents, expertise, criticism and passion. Of course, nothing could have been done without the brave patients who donated their sera — some of them more than once — so that this story could be pursued. The author also thanks the members of his team whose images have been reproduced here. The original work was funded by the US National Institutes of Health (NIH). The writing of this manuscript was supported by The Wellcome Trust, of which the author is a Principal Research Fellow (grant number 073915). The Wellcome Trust Centre for Cell Biology is supported by core grant numbers 077707 and 092076.
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Earnshaw, W. Discovering centromere proteins: from cold white hands to the A, B, C of CENPs. Nat Rev Mol Cell Biol 16, 443–449 (2015). https://doi.org/10.1038/nrm4001
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DOI: https://doi.org/10.1038/nrm4001
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