Centromeres are defined epigenetically and require the presence of the centromere-specific histone H3 variant centromere protein A (CENP-A).
Although DNA sequences are not strictly required for centromere specification, similarities in the organization of centromere DNA suggest that DNA structures contribute to centromere function.
CENP-A nucleosomes contain unique sequence and structural features that allow them to stably mark the centromere and be recognized by kinetochore components.
CENP-A propagation requires specialized deposition factors and tight regulatory control.
The centromere directs the assembly of the kinetochore via the 16-subunit constitutive centromere-associated network (CCAN).
The centromere is the region of the chromosome that directs its segregation in mitosis and meiosis. Although the functional importance of the centromere has been appreciated for more than 130 years, elucidating the molecular features and properties that enable centromeres to orchestrate chromosome segregation is an ongoing challenge. Most eukaryotic centromeres are defined epigenetically and require the presence of nucleosomes containing the histone H3 variant centromere protein A (CENP-A; also known as CENH3). Ongoing work is providing important molecular insights into the central requirements for centromere identity and propagation, and the mechanisms by which centromeres recruit kinetochores to connect to spindle microtubules.
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Holland, A. J. & Cleveland, D. W. Boveri revisited: chromosomal instability, aneuploidy and tumorigenesis. Nat. Rev. Mol. Cell Biol. 10, 478–487 (2009).
Flemming, W. Zellsubstanz, Kern und Zelltheilung (in German) (F. C. W. Vogel, 1882).
Vig, B. K. Sequence of centromere separation: role of centromeric heterochromatin. Genetics 102, 795–806 (1982).
Bernard, P. et al. Requirement of heterochromatin for cohesion at centromeres. Science 294, 2539–2542 (2001).
Nasmyth, K. Segregating sister genomes: the molecular biology of chromosome separation. Science 297, 559–565 (2002).
Kerrebrock, A. W., Moore, D. P., Wu, J. S. & Orr-Weaver, T. L. Mei-S332, a Drosophila protein required for sister-chromatid cohesion, can localize to meiotic centromere regions. Cell 83, 247–256 (1995).
Lindegren, C. C. The genetics of Neurospora III: pure bred stocks and crossing over in N. crassa. Bull. Torrey Bot. Club 60, 133–154 (1933).
Bridges, C. B. & Morgan, T. H. The Third-chromosome Group of Mutant Characters of Drosophila melanogaster (Carnegie Institution of Washington, 1923).
Fukagawa, T. & Earnshaw, W. C. The centromere: chromatin foundation for the kinetochore machinery. Dev. Cell 30, 496–508 (2014).
Darlington, C. D. The external mechanics of chromosomes. I — The scope of enquiry. Proc. R. Soc. B Biol. Sci. 121, 264–319 (1936).
Guerra, M. et al. Neocentrics and holokinetics (holocentrics): chromosomes out of the centromeric rules. Cytogenet. Genome Res. 129, 82–96 (2010).
Pluta, A. F., Mackay, A. M., Ainsztein, A. M., Goldberg, I. G. & Earnshaw, W. C. The centromere: hub of chromosomal activities. Science 270, 1591–1594 (1995).
Clarke, L. & Carbon, J. Isolation of a yeast centromere and construction of functional small circular chromosomes. Nature 287, 504–509 (1980). This paper describes the first cloning of a budding yeast centromere sequence and the sufficiency of this sequence to direct the segregation of exogenous DNA.
Clarke, L. & Carbon, J. Genomic substitutions of centromeres in Saccharomyces cerevisiae. Nature 305, 23–28 (1983).
Carbon, J. & Clarke, L. Structural and functional analysis of a yeast centromere (CEN3). J. Cell Sci. Suppl. 1, 43–58 (1984).
McGrew, J., Diehl, B. & Fitzgerald-Hayes, M. Single base-pair mutations in centromere element III cause aberrant chromosome segregation in Saccharomyces cerevisiae. Mol. Cell. Biol. 6, 530–538 (1986).
Sanyal, K., Baum, M. & Carbon, J. Centromeric DNA sequences in the pathogenic yeast Candida albicans are all different and unique. Proc. Natl Acad. Sci. USA 101, 11374–11379 (2004).
Locke, D. P. et al. Comparative and demographic analysis of orang-utan genomes. Nature 469, 529–533 (2011).
Piras, F. M. et al. Uncoupling of satellite DNA and centromeric function in the genus Equus. PLoS Genet. 6, e1000845 (2010).
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).
Kit, S. Equilibrium sedimentation in density gradients of DNA preparations from animal tissues. J. Mol. Biol. 3, 711–716 (1961).
Malik, H. S. & Henikoff, S. Major evolutionary transitions in centromere complexity. Cell 138, 1067–1082 (2009).
Fishel, B., Amstutz, H., Baum, M., Carbon, J. & Clarke, L. Structural organization and functional analysis of centromeric DNA in the fission yeast Schizosaccharomyces pombe. Mol. Cell. Biol. 8, 754–763 (1988).
Joseph, A., Mitchell, A. R. & Miller, O. J. The organization of the mouse satellite DNA at centromeres. Exp. Cell Res. 183, 494–500 (1989).
Maio, J. J. DNA strand reassociation and polyribonucleotide binding in the African green monkey, Cercopithecus aethiops. J. Mol. Biol. 56, 579–595 (1971).
Rosenberg, H., Singer, M. & Rosenberg, M. Highly reiterated sequences of SIMIANSIMIANSIMIANSIMIANSIMIAN. Science 200, 394–402 (1978).
Manuelidis, L. Chromosomal localization of complex and simple repeated human DNAs. Chromosoma 66, 23–32 (1978).
Manuelidis, L. Complex and simple sequences in human repeated DNAs. Chromosoma 66, 1–21 (1978).
Aldrup-Macdonald, M. E. & Sullivan, B. A. The past, present, and future of human centromere genomics. Genes 5, 33–50 (2014).
Montefalcone, G., Tempesta, S., Rocchi, M. & Archidiacono, N. Centromere repositioning. Genome Res. 9, 1184–1188 (1999). This paper provided the first evidence that centromeres have repositioned over evolutionary history, independently of their surrounding markers.
Rocchi, M., Archidiacono, N., Schempp, W., Capozzi, O. & Stanyon, R. Centromere repositioning in mammals. Heredity 108, 59–67 (2012).
Kasai, F., Garcia, C., Arruga, M. V. & Ferguson-Smith, M. A. Chromosome homology between chicken (Gallus gallus domesticus) and the red-legged partridge (Alectoris rufa); evidence of the occurrence of a neocentromere during evolution. Cytogenet. Genome Res. 102, 326–330 (2003).
Ventura, M. et al. Evolutionary formation of new centromeres in macaque. Science 316, 243–246 (2007).
Kalitsis, P. & Choo, K. H. The evolutionary life cycle of the resilient centromere. Chromosoma 121, 327–340 (2012).
Hahnenberger, K. M., Baum, M. P., Polizzi, C. M., Carbon, J. & Clarke, L. Construction of functional artificial minichromosomes in the fission yeast Schizosaccharomyces pombe. Proc. Natl Acad. Sci. USA 86, 577–581 (1989).
Haaf, T., Warburton, P. E. & Willard, H. F. Integration of human α-satellite DNA into simian chromosomes: centromere protein binding and disruption of normal chromosome segregation. Cell 70, 681–696 (1992).
Harrington, J. J., Van Bokkelen, G., Mays, R. W., Gustashaw, K. & Willard, H. F. Formation of de novo centromeres and construction of first-generation human artificial microchromosomes. Nat. Genet. 15, 345–355 (1997).
Ikeno, M. et al. Construction of YAC-based mammalian artificial chromosomes. Nat. Biotechnol. 16, 431–439 (1998). References 37 and 38 describe the first use of human centromeric DNA to confer mitotic stability on exogenous DNA to generate artificial chromosomes.
Masumoto, H. et al. Assay of centromere function using a human artificial chromosome. Chromosoma 107, 406–416 (1998).
Bergmann, J. H. et al. Epigenetic engineering shows H3K4me2 is required for HJURP targeting and CENP-A assembly on a synthetic human kinetochore. EMBO J. 30, 328–340 (2011).
Ohzeki, J. et al. Breaking the HAC barrier: histone H3K9 acetyl/methyl balance regulates CENP-A assembly. EMBO J. 31, 2391–2402 (2012).
Lechner, J. & Carbon, J. A 240 kd multisubunit protein complex, CBF3, is a major component of the budding yeast centromere. Cell 64, 717–725 (1991).
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).
Muro, Y. et al. Centromere protein B assembles human centromeric alpha-satellite DNA at the 17-bp sequence, CENP-B box. J. Cell Biol. 116, 585–596 (1992).
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). This paper reports the discovery of the first human centromere proteins, CENP-A, CENP-B and CENP-C, as antigens recognized by serum from patients with CREST syndrome.
Haaf, T., Mater, A. G., Wienberg, J. & Ward, D. C. Presence and abundance of CENP-B box sequences in great ape subsets of primate-specific α-satellite DNA. J. Mol. Evol. 41, 487–491 (1995).
Kipling, D. et al. CENP-B binds a novel centromeric sequence in the Asian mouse Mus caroli. Mol. Cell. Biol. 15, 4009–4020 (1995).
Fachinetti, D. et al. A two-step mechanism for epigenetic specification of centromere identity and function. Nat. Cell Biol. 15, 1056–1066 (2013).
Fachinetti, D. et al. DNA sequence-specific binding of CENP-B enhances the fidelity of human centromere function. Dev. Cell 33, 314–327 (2015).
Fujita, R. et al. Stable complex formation of CENP-B with the CENP-A nucleosome. Nucleic Acids Res. 43, 4909–4922 (2015).
Kapoor, M. et al. The cenpB gene is not essential in mice. Chromosoma 107, 570–576 (1998).
Perez-Castro, A. V. et al. Centromeric protein B null mice are viable with no apparent abnormalities. Dev. Biol. 201, 135–143 (1998).
Hudson, D. F. 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).
Voullaire, L. E., Slater, H. R., Petrovic, V. & Choo, K. H. A functional marker centromere with no detectable alpha-satellite, satellite III, or CENP-B protein: activation of a latent centromere? Am. J. Hum. Genet. 52, 1153–1163 (1993). This paper reports the first human neocentromere, demonstrating that centromeres can function in the absence of the human centromeric α-satellite sequences.
Earnshaw, W. C. et al. Molecular cloning of cDNA for CENP-B, the major human centromere autoantigen. J. Cell Biol. 104, 817–829 (1987).
Broccoli, D., Miller, O. J. & Miller, D. A. Relationship of mouse minor satellite DNA to centromere activity. Cytogenet. Cell Genet. 54, 182–186 (1990).
Grimes, B. R., Rhoades, A. A. & Willard, H. F. α-satellite DNA and vector composition influence rates of human artificial chromosome formation. Mol. Ther. 5, 798–805 (2002).
Ohzeki, J., Nakano, M., Okada, T. & Masumoto, H. CENP-B box is required for de novo centromere chromatin assembly on human alphoid DNA. J. Cell Biol. 159, 765–775 (2002).
Palmer, D. K. & Margolis, R. L. Kinetochore components recognized by human autoantibodies are present on mononucleosomes. Mol. Cell. Biol. 5, 173–186 (1985). This paper provided the first indication that centromere components (defined by detection with serum from patients with CREST syndrome and now recognized as CENP-A) are components of chromatin and proposed that such components may be exchanged for canonical histones at the centromere.
Palmer, D. K., O'Day, K. & Margolis, R. L. The centromere specific histone CENP-A is selectively retained in discrete foci in mammalian sperm nuclei. Chromosoma 100, 32–36 (1990).
Palmer, D. K., O'Day, K., Trong, H. L., Charbonneau, H. & Margolis, R. L. Purification of the centromere-specific protein CENP-A and demonstration that it is a distinctive histone. Proc. Natl Acad. Sci. USA 88, 3734–3738 (1991).
Palmer, D. K., O'Day, K., Wener, M. H., Andrews, B. S. & 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, 581–592 (1994). This paper reports the cloning of human CENP-A and defined the importance of its domain containing sequence homology to histone H3.
Buchwitz, B. J., Ahmad, K., Moore, L. L., Roth, M. B. & Henikoff, S. A histone-H3-like protein in C. elegans. Nature 401, 547–548 (1999).
Henikoff, S., Ahmad, K., Platero, J. S. & van Steensel, B. Heterochromatic deposition of centromeric histone H3-like proteins. Proc. Natl Acad. Sci. USA 97, 716–721 (2000).
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).
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).
Vafa, O. & Sullivan, K. F. Chromatin containing CENP-A and α-satellite DNA is a major component of the inner kinetochore plate. Curr. Biol. 7, 897–900 (1997).
Marshall, O. J., Chueh, A. C., Wong, L. H. & Choo, K. H. Neocentromeres: new insights into centromere structure, disease development, and karyotype evolution. Am. J. Hum. Genet. 82, 261–282 (2008).
Earnshaw, W. C. & Migeon, B. R. Three related centromere proteins are absent from the inactive centromere of a stable isodicentric chromosome. Chromosoma 92, 290–296 (1985). This paper provided the first evidence for the epigenetic nature of the centromere, by observing that the inactive centromere of a dicentric chromosome maintained the centromeric DNA structures (as detected by traditional banding techniques) but lacked detectable centromere proteins.
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).
Regnier, V. et al. CENP-A is required for accurate chromosome segregation and sustained kinetochore association of BubR1. Mol. Cell. Biol. 25, 3967–3981 (2005).
Heun, P. et al. Mislocalization of the Drosophila centromere-specific histone CID promotes formation of functional ectopic kinetochores. Dev. Cell 10, 303–315 (2006).
Mendiburo, M. J., Padeken, J., Fulop, S., Schepers, A. & Heun, P. Drosophila CENH3 is sufficient for centromere formation. Science 334, 686–690 (2011).
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).
Logsdon, G. A. et al. Both tails and the centromere targeting domain of CENP-A are required for centromere establishment. J. Cell Biol. 208, 521–531 (2015).
Goutte-Gattat, D. et al. Phosphorylation of the CENP-A amino-terminus in mitotic centromeric chromatin is required for kinetochore function. Proc. Natl Acad. Sci. USA 110, 8579–8584 (2013).
Black, B. E., Brock, M. A., Bedard, S., Woods, V. L. Jr & Cleveland, D. W. An epigenetic mark generated by the incorporation of CENP-A into centromeric nucleosomes. Proc. Natl Acad. Sci. USA 104, 5008–5013 (2007).
Black, B. E. et al. Centromere identity maintained by nucleosomes assembled with histone H3 containing the CENP-A targeting domain. Mol. Cell 25, 309–322 (2007). This study uses chimeric histones containing elements of histone H3 combined with elements of CENP-A to demonstrate that the centromere recruitment of CENP-A is encoded by its first loop and second α-helix, defining the CATD.
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).
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. Nat. Cell Biol. 11, 896–902 (2009). References 80 and 81 report the interaction of CENP-C and CENP-N with CENP-A nucleosomes, providing the first direct physical connections between CENP-A nucleosomes and the proteins of the kinetochore.
Kato, H. et al. A conserved mechanism for centromeric nucleosome recognition by centromere protein CENP-C. Science 340, 1110–1113 (2013).
Guse, A., Carroll, C. W., Moree, B., Fuller, C. J. & Straight, A. F. In vitro centromere and kinetochore assembly on defined chromatin templates. Nature 477, 354–358 (2011).
Westhorpe, F. G., Fuller, C. J. & Straight, A. F. A cell-free CENP-A assembly system defines the chromatin requirements for centromere maintenance. J. Cell Biol. 209, 789–801 (2015).
Chen, Y. et al. The N terminus of the centromere H3-like protein Cse4p performs an essential function distinct from that of the histone fold domain. Mol. Cell. Biol. 20, 7037–7048 (2000).
Van Hooser, A. A. et al. Specification of kinetochore-forming chromatin by the histone H3 variant CENP-A. J. Cell Sci. 114, 3529–3542 (2001).
Folco, H. D. et al. The CENP-A N-tail confers epigenetic stability to centromeres via the CENP-T branch of the CCAN in fission yeast. Curr. Biol. 25, 348–356 (2015).
Black, B. E. et al. Structural determinants for generating centromeric chromatin. Nature 430, 578–582 (2004).
Sekulic, N., Bassett, E. A., Rogers, D. J. & Black, B. E. The structure of (CENP-A–H4)2 reveals physical features that mark centromeres. Nature 467, 347–351 (2010).
Tachiwana, H. et al. Crystal structure of the human centromeric nucleosome containing CENP-A. Nature 476, 232–235 (2011).
Falk, S. J. et al. Chromosomes. CENP-C reshapes and stabilizes CENP-A nucleosomes at the centromere. Science 348, 699–703 (2015).
Dunleavy, E. M., Zhang, W. & Karpen, G. H. Solo or doppio: how many CENP-As make a centromeric nucleosome? Nat. Struct. Mol. Biol. 20, 648–650 (2013).
Panchenko, T. et al. Replacement of histone H3 with CENP-A directs global nucleosome array condensation and loosening of nucleosome superhelical termini. Proc. Natl Acad. Sci. USA 108, 16588–16593 (2011).
Geiss, C. P. et al. CENP-A arrays are more condensed than canonical arrays at low ionic strength. Biophys. J. 106, 875–882 (2014).
Conde e Silva, N. et al. CENP-A-containing nucleosomes: easier disassembly versus exclusive centromeric localization. J. Mol. Biol. 370, 555–573 (2007).
Hasson, D. et al. The octamer is the major form of CENP-A nucleosomes at human centromeres. Nat. Struct. Mol. Biol. 20, 687–695 (2013).
McClintock, B. The behavior in successive nuclear divisions of a chromosome broken at meiosis. Proc. Natl Acad. Sci. USA 25, 405–416 (1939).
Koshland, D., Rutledge, L., Fitzgerald-Hayes, M. & Hartwell, L. H. A genetic analysis of dicentric minichromosomes in Saccharomyces cerevisiae. Cell 48, 801–812 (1987).
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). This paper provides a major advance in understanding the mechanisms that propagate CENP-A nucleosomes, by demonstrating the striking stability of CENP-A at centromeres, its partitioning between replicated sisters during S phase and its new assembly during G1.
Bodor, D. L., Valente, L. P., Mata, J. F., Black, B. E. & Jansen, L. E. Assembly in G1 phase and long-term stability are unique intrinsic features of CENP-A nucleosomes. Mol. Biol. Cell 24, 923–932 (2013).
Dunleavy, E. M., Almouzni, G. & Karpen, G. H. H3.3 is deposited at centromeres in S phase as a placeholder for newly assembled CENP-A in G1 phase. Nucleus 2, 146–157 (2011).
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). References 102 and 103 report the discovery of the CENP-A specific chaperone, HJURP.
Zhou, Z. et al. Structural basis for recognition of centromere histone variant CenH3 by the chaperone Scm3. Nature 472, 234–237 (2011).
Hu, H. et al. Structure of a CENP-A-histone H4 heterodimer in complex with chaperone HJURP. Genes Dev. 25, 901–906 (2011).
Shuaib, M., Ouararhni, K., Dimitrov, S. & Hamiche, A. HJURP binds CENP-A via a highly conserved N-terminal domain and mediates its deposition at centromeres. Proc. Natl Acad. Sci. USA 107, 1349–1354 (2010).
Bassett, E. A. et al. HJURP uses distinct CENP-A surfaces to recognize and to stabilize CENP-A/histone H4 for centromere assembly. Dev. Cell 22, 749–762 (2012).
Sanchez-Pulido, L., Pidoux, A. L., Ponting, C. P. & Allshire, R. C. Common ancestry of the CENP-A chaperones Scm3 and HJURP. Cell 137, 1173–1174 (2009).
Fujita, Y. et al. Priming of centromere for CENP-A recruitment by human hMis18α, hMis18β, and M18BP1. Dev. Cell 12, 17–30 (2007).
Maddox, P. S., Hyndman, F., Monen, J., Oegema, K. & Desai, A. Functional genomics identifies a Myb domain-containing protein family required for assembly of CENP-A chromatin. J. Cell Biol. 176, 757–763 (2007).
Hayashi, T. et al. Mis16 and Mis18 are required for CENP-A loading and histone deacetylation at centromeres. Cell 118, 715–729 (2004). References 109–111 report the discovery of the components of the MIS18 complex, which is crucial for CENP-A deposition.
Erhardt, S. et al. Genome-wide analysis reveals a cell cycle-dependent mechanism controlling centromere propagation. J. Cell Biol. 183, 805–818 (2008).
Chen, C. C. et al. CAL1 is the Drosophila CENP-A assembly factor. J. Cell Biol. 204, 313–329 (2014).
Dambacher, S. et al. CENP-C facilitates the recruitment of M18BP1 to centromeric chromatin. Nucleus 3, 101–110 (2012).
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).
McKinley, K. L. & Cheeseman, I. M. Polo-like kinase 1 licenses CENP-A deposition at centromeres. Cell 158, 397–411 (2014).
Tachiwana, H. et al. HJURP involvement in de novo CenH3CENP-A and CENP-C recruitment. Cell Rep. 11, 22–32 (2015).
Perpelescu, M., Nozaki, N., Obuse, C., Yang, H. & Yoda, K. Active establishment of centromeric CENP-A chromatin by RSF complex. J. Cell Biol. 185, 397–407 (2009).
Lagana, A. et al. A small GTPase molecular switch regulates epigenetic centromere maintenance by stabilizing newly incorporated CENP-A. Nat. Cell Biol. 12, 1186–1193 (2010).
Akiyoshi, B. & Gull, K. Discovery of unconventional kinetochores in kinetoplastids. Cell 156, 1247–1258 (2014).
Drinnenberg, I. A., deYoung, D., Henikoff, S. & Malik, H. S. Recurrent loss of CenH3 is associated with independent transitions to holocentricity in insects. eLife 3, e03676 (2014).
Bodor, D. L. et al. The quantitative architecture of centromeric chromatin. eLife 3, e02137 (2014).
Lima-de-Faria, A. Genetics, origin, and evolution of kinetochores. Hereditas 35, 422–444 (1949).
Peters, A. H. et al. Partitioning and plasticity of repressive histone methylation states in mammalian chromatin. Mol. Cell 12, 1577–1589 (2003).
Rice, J. C. et al. Histone methyltransferases direct different degrees of methylation to define distinct chromatin domains. Mol. Cell 12, 1591–1598 (2003).
Shang, W. H. et al. Chromosome engineering allows the efficient isolation of vertebrate neocentromeres. Dev. Cell 24, 635–648 (2013).
Alonso, A., Hasson, D., Cheung, F. & Warburton, P. E. A paucity of heterochromatin at functional human neocentromeres. Epigenetics Chromatin 3, 6 (2010).
Blower, M. D., Sullivan, B. A. & Karpen, G. H. Conserved organization of centromeric chromatin in flies and humans. Dev. Cell 2, 319–330 (2002).
Sullivan, B. A. & Karpen, G. H. Centromeric chromatin exhibits a histone modification pattern that is distinct from both euchromatin and heterochromatin. Nat. Struct. Mol. Biol. 11, 1076–1083 (2004). This paper provides clear evidence for two distinct chromatin signatures within the centromere, demonstrating that the core centromere contains euchromatic marks, including H3K4me2, and lacks the heterochromatic marks of the pericentromere.
Ribeiro, S. A. et al. A super-resolution map of the vertebrate kinetochore. Proc. Natl Acad. Sci. USA 107, 10484–10489 (2010).
Nakano, M. et al. Inactivation of a human kinetochore by specific targeting of chromatin modifiers. Dev. Cell 14, 507–522 (2008).
Folco, H. D., Pidoux, A. L., Urano, T. & Allshire, R. C. Heterochromatin and RNAi are required to establish CENP-A chromatin at centromeres. Science 319, 94–97 (2008).
Choi, E. S. et al. Identification of noncoding transcripts from within CENP-A chromatin at fission yeast centromeres. J. Biol. Chem. 286, 23600–23607 (2011).
Rosic, S., Kohler, F. & Erhardt, S. Repetitive centromeric satellite RNA is essential for kinetochore formation and cell division. J. Cell Biol. 207, 335–349 (2014).
Chan, F. L. et al. Active transcription and essential role of RNA polymerase II at the centromere during mitosis. Proc. Natl Acad. Sci. USA 109, 1979–1984 (2012).
Liu, H. et al. Mitotic transcription installs Sgo1 at centromeres to coordinate chromosome segregation. Mol. Cell 59, 426–436 (2015).
Quenet, D. & Dalal, Y. A long non-coding RNA is required for targeting centromeric protein A to the human centromere. eLife 3, e03254 (2014).
Hill, A. & Bloom, K. Genetic manipulation of centromere function. Mol. Cell. Biol. 7, 2397–2405 (1987).
Okada, M., Okawa, K., Isobe, T. & Fukagawa, T. CENP-H-containing complex facilitates centromere deposition of CENP-A in cooperation with FACT and CHD1. Mol. Biol. Cell 20, 3986–3995 (2009).
Furuyama, T., Dalal, Y. & Henikoff, S. Chaperone-mediated assembly of centromeric chromatin in vitro. Proc. Natl Acad. Sci. USA 103, 6172–6177 (2006).
Chen, C. C. et al. Establishment of centromeric chromatin by the CENP-A assembly factor CAL1 requires FACT-mediated transcription. Dev. Cell 34, 73–84 (2015).
Kim, I. S. et al. Roles of Mis18α in epigenetic regulation of centromeric chromatin and CENP-A loading. Mol. Cell 46, 260–273 (2012).
Wang, J. et al. Mitotic regulator Mis18β interacts with and specifies the centromeric assembly of molecular chaperone holliday junction recognition protein (HJURP). J. Biol. Chem. 289, 8326–8336 (2014).
Perpelescu, M. et al. HJURP is involved in the expansion of centromeric chromatin. Mol. Biol. Cell 26, 2742–2754 (2015).
Schuh, M., Lehner, C. F. & Heidmann, S. Incorporation of Drosophila CID/CENP-A and CENP-C into centromeres during early embryonic anaphase. Curr. Biol. 17, 237–243 (2007).
Mellone, B. G. et al. Assembly of Drosophila centromeric chromatin proteins during mitosis. PLoS Genet. 7, e1002068 (2011).
Nechemia-Arbely, Y., Fachinetti, D. & Cleveland, D. W. Replicating centromeric chromatin: spatial and temporal control of CENP-A assembly. Exp. Cell Res. 318, 1353–1360 (2012).
Silva, M. C. et al. Cdk activity couples epigenetic centromere inheritance to cell cycle progression. Dev. Cell 22, 52–63 (2012).
Muller, S. et al. Phosphorylation and DNA binding of HJURP determine its centromeric recruitment and function in CenH3CENP-A loading. Cell Rep. 8, 190–203 (2014).
Yu, Z. et al. Dynamic phosphorylation of CENP-A at Ser68 orchestrates its cell-cycle-dependent deposition at centromeres. Dev. Cell 32, 68–81 (2015).
Bell, S. P. & Dutta, A. DNA replication in eukaryotic cells. Annu. Rev. Biochem. 71, 333–374 (2002).
Collins, K. A., Furuyama, S. & Biggins, S. Proteolysis contributes to the exclusive centromere localization of the yeast Cse4/CENP-A histone H3 variant. Curr. Biol. 14, 1968–1972 (2004).
Deyter, G. M. & Biggins, S. The FACT complex interacts with the E3 ubiquitin ligase Psh1 to prevent ectopic localization of CENP-A. Genes Dev. 28, 1815–1826 (2014).
Kitagawa, T., Ishii, K., Takeda, K. & Matsumoto, T. The 19S proteasome subunit Rpt3 regulates distribution of CENP-A by associating with centromeric chromatin. Nat. Commun. 5, 3597 (2014).
Spence, J. M. et al. Co-localization of centromere activity, proteins and topoisomerase II within a subdomain of the major human X α-satellite array. EMBO J. 21, 5269–5280 (2002).
Zeng, K. et al. Localisation of centromeric proteins to a fraction of mouse minor satellite DNA on a mini-chromosome in human, mouse and chicken cells. Chromosoma 113, 84–91 (2004).
Sullivan, L. L., Boivin, C. D., Mravinac, B., Song, I. Y. & Sullivan, B. A. Genomic size of CENP-A domain is proportional to total alpha satellite array size at human centromeres and expands in cancer cells. Chromosome Res. 19, 457–470 (2011).
Lo, A. W. et al. A novel chromatin immunoprecipitation and array (CIA) analysis identifies a 460-kb CENP-A-binding neocentromere DNA. Genome Res. 11, 448–457 (2001).
Gascoigne, K. E. et al. Induced ectopic kinetochore assembly bypasses the requirement for CENP-A nucleosomes. Cell 145, 410–422 (2011).
Schittenhelm, R. B., Althoff, F., Heidmann, S. & Lehner, C. F. Detrimental incorporation of excess Cenp-A/Cid and Cenp-C into Drosophila centromeres is prevented by limiting amounts of the bridging factor Cal1. J. Cell Sci. 123, 3768–3779 (2010).
Cheeseman, I. M. & Desai, A. Molecular architecture of the kinetochore–microtubule interface. Nat. Rev. Mol. Cell Biol. 9, 33–46 (2008).
Foltz, D. R. et al. The human CENP-A centromeric nucleosome-associated complex. Nat. Cell Biol. 8, 458–469 (2006).
Okada, M. et al. The CENP-H-I complex is required for the efficient incorporation of newly synthesized CENP-A into centromeres. Nat. Cell Biol. 8, 446–457 (2006).
Izuta, H. et al. Comprehensive analysis of the ICEN (Interphase Centromere Complex) components enriched in the CENP-A chromatin of human cells. Genes Cells 11, 673–684 (2006). References 162–164 report the identification of the majority of the components of the human CCAN by mass spectrometry analysis.
Amano, M. et al. The CENP-S complex is essential for the stable assembly of outer kinetochore structure. J. Cell Biol. 186, 173–182 (2009).
Saitoh, H. et al. CENP-C, an autoantigen in scleroderma, is a component of the human inner kinetochore plate. Cell 70, 115–125 (1992).
Nishihashi, A. et al. CENP-I is essential for centromere function in vertebrate cells. Dev. Cell 2, 463–476 (2002).
Takahashi, K., Yamada, H. & Yanagida, M. Fission yeast minichromosome loss mutants mis cause lethal aneuploidy and replication abnormality. Mol. Biol. Cell 5, 1145–1158 (1994).
Minoshima, Y. et al. The constitutive centromere component CENP-50 is required for recovery from spindle damage. Mol. Cell. Biol. 25, 10315–10328 (2005).
Sugata, N., Munekata, E. & Todokoro, K. Characterization of a novel kinetochore protein, CENP-H. J. Biol. Chem. 274, 27343–27346 (1999).
Hinshaw, S. M. & Harrison, S. C. An Iml3-Chl4 heterodimer links the core centromere to factors required for accurate chromosome segregation. Cell Rep. 5, 29–36 (2013).
Basilico, F. et al. The pseudo GTPase CENP-M drives human kinetochore assembly. eLife 3, e02978 (2014).
Hornung, P. et al. A cooperative mechanism drives budding yeast kinetochore assembly downstream of CENP-A. J. Cell Biol. 206, 509–524 (2014).
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).
Nishino, T. et al. CENP-T-W-S-X forms a unique centromeric chromatin structure with a histone-like fold. Cell 148, 487–501 (2012). This work reveals that the histone-fold domains of the kinetochore proteins CENP-T, -W, -S and -X form a complex that resembles the arrangement of histones in a nucleosome.
Westermann, S. & Schleiffer, A. Family matters: structural and functional conservation of centromere-associated proteins from yeast to humans. Trends Cell Biol. 23, 260–269 (2013).
Schleiffer, A. et al. CENP-T proteins are conserved centromere receptors of the Ndc80 complex. Nat. Cell Biol. 14, 604–613 (2012). In this paper, the authors identify the long-elusive yeast homologues of numerous CCAN components.
Ortiz, J., Stemmann, O., Rank, S. & Lechner, J. A putative protein complex consisting of Ctf19, Mcm21, and Okp1 represents a missing link in the budding yeast kinetochore. Genes Dev. 13, 1140–1155 (1999).
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).
Kagawa, N. et al. The CENP-O complex requirement varies among different cell types. Chromosome Res. 22, 293–303 (2014).
Klare, K. et al. CENP-C is a blueprint for constitutive centromere-associated network assembly within human kinetochores. J. Cell Biol. 210, 11–22 (2015).
Nagpal, H. et al. Dynamic changes in the CCAN organization through CENP-C during cell-cycle progression. Mol. Biol. Cell 26, 3768–3776 (2015).
McKinley, K. L. et al. The CENP-L-N complex forms a critical node in an integrated meshwork of interactions at the centromere-kinetochore interface. Mol. Cell http://dx.doi.org/10.1016/j.molcel.2015.10.027 (2015).
Kwon, M. S., Hori, T., Okada, M. & Fukagawa, T. CENP-C is involved in chromosome segregation, mitotic checkpoint function, and kinetochore assembly. Mol. Biol. Cell 18, 2155–2168 (2007).
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).
Hori, T. et al. CCAN makes multiple contacts with centromeric DNA to provide distinct pathways to the outer kinetochore. Cell 135, 1039–1052 (2008).
Takeuchi, K. et al. The centromeric nucleosome-like CENP-T-W-S-X complex induces positive supercoils into DNA. Nucleic Acids Res. 42, 1644–1655 (2014).
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).
Przewloka, M. R. et al. CENP-C is a structural platform for kinetochore assembly. Curr. Biol. 21, 399–405 (2011).
Malvezzi, F. et al. A structural basis for kinetochore recruitment of the Ndc80 complex via two distinct centromere receptors. EMBO J. 32, 409–423 (2013).
Nishino, T. et al. CENP-T provides a structural platform for outer kinetochore assembly. EMBO J. 32, 424–436 (2013).
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).
Kim, S. & Yu, H. Multiple assembly mechanisms anchor the KMN spindle checkpoint platform at human mitotic kinetochores. J. Cell Biol. 208, 181–196 (2015).
Gascoigne, K. E. & Cheeseman, I. M. CDK-dependent phosphorylation and nuclear exclusion coordinately control kinetochore assembly state. J. Cell Biol. 201, 23–32 (2013).
Rago, F., Gascoigne, K. E. & Cheeseman, I. M. Distinct organization and regulation of the outer kinetochore KMN network downstream of CENP-C and CENP-T. Curr. Biol. 25, 671–677 (2015).
Suzuki, A., Badger, B. L., Wan, X., DeLuca, J. G. & Salmon, E. D. The architecture of CCAN proteins creates a structural integrity to resist spindle forces and achieve proper intrakinetochore stretch. Dev. Cell 30, 717–730 (2014).
Amaro, A. C. et al. Molecular control of kinetochore-microtubule dynamics and chromosome oscillations. Nat. Cell Biol. 12, 319–329 (2010).
Bancroft, J., Auckland, P., Samora, C. P. & McAinsh, A. D. Chromosome congression is promoted by CENP-Q- and CENP-E-dependent pathways. J. Cell Sci. 128, 171–184 (2015).
Steiner, N. C. & Clarke, L. A novel epigenetic effect can alter centromere function in fission yeast. Cell 79, 865–874 (1994).
Higgins, A. W., Gustashaw, K. M. & Willard, H. F. Engineered human dicentric chromosomes show centromere plasticity. Chromosome Res. 13, 745–762 (2005).
Sato, H., Masuda, F., Takayama, Y., Takahashi, K. & Saitoh, S. Epigenetic inactivation and subsequent heterochromatinization of a centromere stabilize dicentric chromosomes. Curr. Biol. 22, 658–667 (2012).
Sullivan, B. A. & Schwartz, S. Identification of centromeric antigens in dicentric Robertsonian translocations: CENP-C and CENP-E are necessary components of functional centromeres. Hum. Mol. Genet. 4, 2189–2197 (1995).
Lange, J. et al. Isodicentric Y chromosomes and sex disorders as byproducts of homologous recombination that maintains palindromes. Cell 138, 855–869 (2009).
Tyler-Smith, C. et al. Transmission of a fully functional human neocentromere through three generations. Am. J. Hum. Genet. 64, 1440–1444 (1999).
Amor, D. J. et al. Human centromere repositioning “in progress”. Proc. Natl Acad. Sci. USA 101, 6542–6547 (2004). This paper reports a chromosome from a human patient containing a neocentromere that lacks α-satellite DNA, as well as a second site containing canonical centromeric α-satellite DNA without centromere function, providing a model for how centromere repositioning over evolutionary time may occur.
Williams, B. C., Murphy, T. D., Goldberg, M. L. & Karpen, G. H. Neocentromere activity of structurally acentric mini-chromosomes in Drosophila. Nat. Genet. 18, 30–37 (1998).
Platero, J. S., Ahmad, K. & Henikoff, S. A distal heterochromatic block displays centromeric activity when detached from a natural centromere. Mol. Cell 4, 995–1004 (1999).
Ketel, C. et al. Neocentromeres form efficiently at multiple possible loci in Candida albicans. PLoS Genet. 5, e1000400 (2009).
Ishii, K. et al. Heterochromatin integrity affects chromosome reorganization after centromere dysfunction. Science 321, 1088–1091 (2008).
Milks, K. J., Moree, B. & Straight, A. F. Dissection of CENP-C-directed centromere and kinetochore assembly. Mol. Biol. Cell 20, 4246–4255 (2009).
Dunleavy, E. M. et al. The cell cycle timing of centromeric chromatin assembly in Drosophila meiosis is distinct from mitosis yet requires CAL1 and CENP-C. PLoS Biol. 10, e1001460 (2012).
Raychaudhuri, N. et al. Transgenerational propagation and quantitative maintenance of paternal centromeres depends on Cid/Cenp-A presence in Drosophila sperm. PLoS Biol. 10, e1001434 (2012).
Gassmann, R. et al. An inverse relationship to germline transcription defines centromeric chromatin in C. elegans. Nature 484, 534–537 (2012).
Schubert, V., Lermontova, I. & Schubert, I. Loading of the centromeric histone H3 variant during meiosis — how does it differ from mitosis? Chromosoma 123, 491–497 (2014).
Kline-Smith, S. L., Khodjakov, A., Hergert, P. & Walczak, C. E. Depletion of centromeric MCAK leads to chromosome congression and segregation defects due to improper kinetochore attachments. Mol. Biol. Cell 15, 1146–1159 (2004).
Schueler, M. G., Higgins, A. W., Rudd, M. K., Gustashaw, K. & Willard, H. F. Genomic and genetic definition of a functional human centromere. Science 294, 109–115 (2001).
Zasadzinska, E., Barnhart-Dailey, M. C., Kuich, P. H. & Foltz, D. R. Dimerization of the CENP-A assembly factor HJURP is required for centromeric nucleosome deposition. EMBO J. 32, 2113–2124 (2013).
Niikura, Y. et al. CENP-A K124 ubiquitylation is required for CENP-A deposition at the centromere. Dev. Cell 32, 589–603 (2015).
Bui, M. et al. Cell-cycle-dependent structural transitions in the human CENP-A nucleosome in vivo. Cell 150, 317–326 (2012).
Zeitlin, S. G., Shelby, R. D. & Sullivan, K. F. CENP-A is phosphorylated by Aurora B kinase and plays an unexpected role in completion of cytokinesis. J. Cell Biol. 155, 1147–1157 (2001).
Bailey, A. O. et al. Posttranslational modification of CENP-A influences the conformation of centromeric chromatin. Proc. Natl Acad. Sci. USA 110, 11827–11832 (2013).
Hori, T. et al. Histone H4 Lys 20 monomethylation of the CENP-A nucleosome is essential for kinetochore assembly. Dev. Cell 29, 740–749 (2014).
The authors apologize to those colleagues whose work they were unable to describe owing to space constraints. They thank members of the Cheeseman laboratory for critical reading of the manuscript and helpful discussions, Bill Earnshaw for directing them to Cyril Darlington's description of the form and function of the centromere, and Conly Rieder, Alexey Khodjakov and Elaine Dunleavy for generously sharing micrographs. Work in the Cheeseman laboratory is supported by a Scholar award to I.M.C. from the Leukemia & Lymphoma Society, a grant from the U.S. National Institutes of Health/National Institute of General Medical Sciences to I.M.C. (GM088313), and a Research Scholar Grant to I.M.C. (121776) from the American Cancer Society.
The authors declare no competing financial interests.
- Meiotic drive
Preferential transmission of a genetic element during meiosis, such that it is represented in more than 50% of the gametes of a heterozygote.
- Evolutionary new centromeres
(ENCs). Centromeres at a different site from the centromere of the chromosome ancestor, for which the movement of the centromere cannot be parsimoniously explained by a simple chromosome rearrangement.
Regions of chromosomes that have the functional characteristics of a centromere, but occur at a site distinct from the site of centromere formation for the chromosome in most organisms of the species, and lack canonical centromere DNA sequences.
- Human artificial chromosomes
(HACs). Units of exogenous DNA that segregate autonomously in human cells.
- Histone chaperone
A protein that binds to histones to facilitate nucleosome assembly.
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McKinley, K., Cheeseman, I. The molecular basis for centromere identity and function. Nat Rev Mol Cell Biol 17, 16–29 (2016). https://doi.org/10.1038/nrm.2015.5
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