Centromere proteins and chromosome inheritance: A complex affair

Current Opinion in Genetics and Development

Volumn 9, Issue 2, 1999, Pages 206-217

  Dobie, Kenneth W ^a   Hari, Kumar L ^a   Maggert, Keith A ^a   Karpen, Gary H ^a  

^a SALK INSTITUTE FOR BIOLOGICAL STUDIES   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CHROMOSOME PROTEIN; STRUCTURAL PROTEIN;

CELL CYCLE; CENTROMERE; INHERITANCE; MEIOSIS; MICROTUBULE; MITOSIS; PRIORITY JOURNAL; REVIEW;

EID: 0033118347     PISSN: 0959437X     EISSN: None     Source Type: Journal    
DOI: 10.1016/S0959-437X(99)80031-8     Document Type: Article

References (126)

1. Hook E: The impact of aneuploidy upon public health: Mortality and morbidity associated with human chromosomal abnormalities. In Aneuploidy: Etiology & Mechanisms, vol. 36. Edited by Dellarco V, Voytek P, Hollaender A. New York: Plenum press; 1985:7-33.
2. Jacobs PA, Hassold TJ: The origin of numerical chromosome abnormalities. Adv Genet 1995, 33:101-133.
3. Mitelman F: Catalog of Chromosome Aberrations in Cancer, 5th Edition. New York: Wiley; 1994.
4. Willard HF: Human artificial chromosomes coming into focus. Nat Biotechnol 1998, 16:415-416.
5. Murphy TD, Karpen GH: Centromeres take flight: Alpha satellite and the quest for the human centromere. Cell 1998, 93:317-320.
6. Clarke L: Centromeres: Proteins, protein complexes, and repeated domains at centromeres of simple eukaryotes. Curr Opin Genet Dev 1998, 8:212-218.
7. Karpen GH, Allshire R: The case for epigenetic effects on centromere identity and function. Trends Genet 1997, 13:489-496.
8. Wiens G R, Sorger PK: Centromeric chromatin and epigenetic effects in kinetochore assembly. Cell 1998, 93:313-316.
9. Espelin CW, Kaplan KB, Sorger PK: Probing the architecture of a simple kinetochore using DNA-protein crosslinking. J Cell Biol 1997, 139:1383-1396.
10. Meluh PB, Koshland D: Budding yeast centromere composition and assembly as revealed by in vivo cross-linking. Genes Dev 1997, 11:3401-3412.
11. Meluh PB, Yang P, Glowczewski L, Koshland D, Smith MM: Cse4p is a component of the core centromere of Saccharomyces cerevisiae. Cell 1998, 94:607-613. The link between baker's yeast and more complex organisms is strengthened by determining that Cse4p, the S. cerevisiae CENP-A homolog, is found at the S. cerevisiae centromere. By assaying the nuclease sensitivity of CEN DNA in cse4 mutant cells and performing chromatin immunoprecipitation and immunolocalization studies, these workers demonstrate that Cse4p interacts with CEN DNA in vivo. A model for the interaction of S. cerevisiae structural proteins with CEN DNA is also presented.
12. Sullivan KF: A moveable feast: The centromere-kinetochore complex in cell division. In Dynamics of Cell Division, vol 20. Edited by Endow SA, Glover DM. Oxford: Oxford University Press; 1998:124-163.
13. Ekwall K, Olsson T, Turner BM, Cranston G, Allshire RC: Transient inhibition of histone deacetylation alters the structural and functional imprint at fisson yeast centromeres. Cell 1997, 91:1021-1032. The treatment of S. pombe cells with TSA, a histone deacetylase inhibitor, causes slow growth, impairs centromeric gene silencing and induces chromosome missegregation. Remarkably, these phenotypes are propagated through >80 generations in the absence of the drug, with a 'flip-back' frequency of just 2% per cell division. TSA exposure also disrupts the centromeric localization of the chromodomain protein Swi6p; however, the protein apparently relocalizes upon drug removal. Nevertheless, histone acetylation may be an epigenetic component that helps define the S. pombe centromere.
14. Csink AK, Henikoff S: Something from nothing: The evolution and utility of satellite repeats. Trends Genet 1998, 14:200-204.
15. Pluta AF, Mackay AM, Ainsztein AM, Goldberg IG, Earnshaw WC: The centromere: Hub of chromosomal activities. Science 1995, 270:1591-1594.
16. He D, Zeng C, Woods K, Zhong L, Turner D, Busch RK, Brinkley BR, Busch H: CENP-G: A new centromeric protein that is associated with the alpha-1 satellite DNA subfamily. Chromosoma 1998, 107:189-197. A new constitutive centromere protein was identified from a patient with watermelon stomach disease (gastric antral vascular ectasia [GAVE]). CENP-G migrates through one-and two-dimensional gels distinct from CENP-A, CENP-B and CENP-C, and appears to be associated with the nuclear matrix during interphase. Immunofluorescence using the GAVE serum shows a double-dot staining pattern on metaphase chromosomes from HeLa and Indian Muntjac cells, and the antiserum labels the inner kinetochore plate of Indian Muntjac metaphase chromosomes in immuno-EM studies.
17. Warburton PE, Cooke CA, Bourassa S, Vafa O, Sullivan BA, Stetten G, Gimelli G, Warburton D, Tyler-Smith C, Sullivan KF et al.: Immunolocalization of CENP-A suggests a distinct nucleosome structure at the inner kinetochore plate of active centromeres. Curr Biol 1997, 7:901-904.
18. Vafa O, Sullivan KF: Chromatin containing CENP-A and alpha-satellite DNA is a major component of the inner kinetochore plate. Curr Biol 1997, 7:897-900.
19. Yoda K, Ando S, Okuda A, Kikuchi A, Okazaki T: In vitro assembly of the CENP-B/alpha-satellite DNA/core histone complex: CENP-B causes nucleosome positioning. Genes Cells 1998, 3:533-548.
20. Perez-Castro AV, Shamanski FL, Meneses JJ, Lovato TL, Vogel KG, Moyzis RK, Pedersen R: Centromeric protein B null mice are viable with no apparent abnormalities. Dev Biol 1998, 201:135-143.
21. Hudson DF, Fowler KJ, Earle E, Saffery R, Kalitsis P, Trowell H, Hill J, Wreford NG, de Kretser DM, Cancilla MR et al.: Centromere protein B null mice are mitotically and meiotically normal but have lower body and testis weights. J Cell Biol 1998, 141:309-319.
22. -/- animals include embryonic lethality, nuclear morphology defects and micronuclei formation. In addition, mitotic chromosomes appear hypercondensed and their segregation is impaired.
23. Fukagawa T, Brown WR: Efficient conditional mutation of the vertebrate CENP-C gene. Hum Mol Genet 1997, 6:2301-2308.
24. Tomkiel J, Cooke CA, Saitoh H, Bernat RL, Earnshaw WC: CENP-C is required for maintaining proper kinetochore size and for a timely transition to anaphase. J Cell Biol 1994, 125:531-545.
25. Pluta AF, Earnshaw WC, Goldberg IG: 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 1998, 111:2029-2041.
26. Pluta AF, Earnshaw WC: Specific interaction between human kinetochore protein CENP-C and a nucleolar transcriptional regulator. J Biol Chem 1996, 271:18767-18774.
27. 1 phase. Proc Natl Acad Sci USA 1996, 93:10234-10239.
28. Takahashi K, Yamada H, Yanagida M: Fission yeast minichromosome loss mutants mis cause lethal aneuploidy and replication abnormality. Mol Biol Cell 1994, 5:1145-1158.
29. Allshire RC, Nimmo ER, Ekwall K, Javerzat JP, Cranston G: Mutations derepressing silent centromeric domains in fission yeast disrupt chromosome segregation. Genes Dev 1995, 9:218-233.
30. Sun X, Wahlstrom J, Karpen GH: Molecular structure of a functional Drosophila centromere. Cell 1997, 91:1007-1019.
31. Cavalli G, Paro R: Chromo-domain proteins: Linking chromatin structure to epigenetic regulation. Curr Opin Cell Biol 1998, 10:354-360.
32. Ekwall K, Javerzat JP, Lorentz A, Schmidt H, Cranston G, Allshire R: The chromodomain protein Swi6: A key component at fission yeast centromeres. Science 1995, 269:1429-1431.
33. Kellum R, Raff JW, Alberts BM: Heterochromatin protein 1 distribution during development and during the cell cycle in Drosophila embryos. J Cell Sci 1995, 108:1407-1418.
34. Wreggett KA, Hill F, James PS, Hutchings A, Butcher GW, Singh PB: A mammalian homologue of Drosophila heterochromatin protein 1 (HP1) is a component of constitutive heterochromatin. Cytogenet Cell Genet 1994, 66:99-103.
35. Wang G, Horsley D, Ma A, Otte AP, Hutchings A, Butcher GW, Singh PB: M33, a mammalian homologue of Drosophila polycomb localises to euchromatin within interphase nuclei but is enriched within the centromeric heterochromatin of metaphase chromosomes. Cytogenet Cell Genet 1997, 78:50-55.
36. Ivanova AV, Bonaduce MJ, Ivanov SV, Klar AJ: The chromo and SET domains of the Clr4 protein are essential for silencing in fission yeast. Nat Genet 1998, 19:192-195.
37. Kellum R, Alberts BM: Heterochromatin protein 1 is required for correct chromosome segregation in Drosophila embryos. J Cell Sci 1995, 108:1419-1431.
38. Rieder CL, Salmon ED: The vertebrate cell kinetochore and its roles during mitosis. Trends Cell Biol 1998, 8:310-318. This outstanding review summarizes a current model for the roles of the kinetochore in chromosome movements throughout cell division. The proposition that the kinetochore MTs exist in moving and neutral states is used to explain MT capture, chromosome congression, and metaphase plate oscillations. The review is a thorough treatment of data from many organisms that seminally contributes to our understanding of how chromosomes are directed in their motions.
39. Hirano T: SMC protein complexes and higher-order chromosome dynamics. Curr Opin Cell Biol 1998, 10:317-322.
40. Jessberger R, Frei C, Gasser SM: Chromosome dynamics: The SMC protein family. Curr Opin Genet Dev 1998, 8:254-259.
41. Moore DP, Orr-Weaver TL: Chromosome segregation during meiosis: Building an unambivalent bivalent. Curr Top Dev Biol 1998, 37:263-299.
42. Pfarr CM, Coue M, Grissom PM, Hays TS, Porter ME, McIntosh JR: Cytoplasmic dynein is localized to kinetochores during mitosis. Nature 1990, 345:263-265.
43. Steuer ER, Wordeman L, Schroer TA, Sheetz MP: Localization of cytoplasmic dynein to mitotic spindles and kinetochores. Nature 1990, 345:266-268.
44. Echeverri CJ, Paschal BM, Vaughan KT, Vallee RB: Molecular characterization of the 50-kD subunit of dynactin reveals function for the complex in chromosome alignment and spindle organization during mitosis. J Cell Biol 1996, 132:617-633.
45. Vig BK: Recognition of mammalian centromeres by anti-dynein and anti-dynactin components. Mutagenesis 1998, 13:391-396.
46. Starr DA, Williams BC, Hays TS, Goldberg ML: ZW10 helps recruit dynactin and dynein to the kinetochore. J Cell Biol 1998, 142:763-774. In this report, Starr et al. report on a cellular phenotype of I(1)zw10 mutants of Drosophila. The authors show that dynein and dynamitin (the p50 subunit of dynactin) are found at active kinetochores in spermatocytes. This localization depends on ZW10, and on ROD (shown previously by Karess and Glover, J Cell Biol 1989, 109:2951-2961 ). The directness of the interaction is supported by a two-hybrid interaction between dynamitin and hZW10. In an intriguing experiment, dynein immunolocalization seems to be moderated by tension, suggesting that the 'tension sensing' mechanism could act upstream of resident motors and MT-binding proteins.
47. Williams BC, Gatti M, Goldberg ML: Bipolar spindle attachments affect redistributions of ZW10, a Drosophila centromere/kinetochore component required for accurate chromosome segregation. J Cell Biol 1996, 134:1127-1140.
48. Williams BC, Karr TL, Montgomery JM, Goldberg ML: The Drosophila I(1)zw10 gene product, required for accurate mitotic chromosome segregation, is redistributed at anaphase onset. J Cell Biol 1992, 118:759-773.
49. Cooke CA, Schaar B, Yen TJ, Earnshaw WC: Localization of CENP-E in the fibrous corona and outer plate of mammalian kinetochores from prometaphase through anaphase. Chromosoma 1997, 106:446-455.
50. Faulkner NE, Vig B, Echeverri CJ, Wordeman L, Vallee RB: Localization of motor-related proteins and associated complexes to active, but not inactive, centromeres. Hum Mol Genet 1998, 7:671-677.
51. Yao X, Anderson KL, Cleveland DW: The microtubule-dependent motor centromere-associated protein E (CENP-E) is an integral component of kinetochore corona fibers that link centromeres to spindle microtubules. J Cell Biol 1997, 139:435-447.
52. Schaar BT, Chan GK, Maddox P, Salmon ED, Yen TJ: CENP-E function at kinetochores is essential for chromosome alignment. J Cell Biol 1997, 139:1373-1382. One of two studies (see also [53•]) showing the role of CENP-E in bipolar spindle formation and chromosome congression. Human cells immunodepleted for CENP-E are capable of entering mitosis but arrest between the formation of monopolar spindles and congression to the metaphase plate. The authors posit two periods of CENP-E activity: the capture of MTs from the distant pole by a monopolar kinetochore complex, making a bipolar spindle, and later during chromosome movement.
53. Wood KW, Sakowicz R, Goldstein LS, Cleveland DW: CENP-E is a plus end-directed kinetochore motor required for metaphase chromosome alignment. Cell 1997, 91:357-366. One of two studies (see also [52•]) describing the effect of CENP-E immunodepletion on mitosis. Xenopus CENP-E was identified and characterized as a plus-end-directed motor in vitro. Antibodies were raised and used to immunodeplete egg extracts. The authors note a moderate increase in monopolar spindles in immunodepleted extracts but a dramatic increase in misaligned bipolar spindles: this is due to failure to form proper bipolar spindles or the inability to maintain them once chromosomes begin to congress. The authors suggest that CENP-E works in two ways at the kinetochore prior to anaphase. On one kinetochore face, CENP-E acts as a motor, moving the chromosomes to the end of a MT. At the other face of the kinetochore, CENP-E acts as a MT-binding protein, always tracking the end of the dynamically unstable MT, behaving in essence as a minus-end-directed motor.
54. Dujardin D, Wacker UI, Moreau A, Schroer TA, Rickard JE, De Mey JR: Evidence for a role of CLIP-170 in the establishment of metaphase chromosome alignment. J Cell Biol 1998, 141:849-862.
55. Doe CL, Wang G, Chow C, Fricker MD, Singh PB, Mellor EJ: The fission yeast chromo domain encoding gene chp1(+) is required for chromosome segregation and shows a genetic interaction with alpha-tubulin. Nucleic Acids Res 1998, 26:4222-4229.
56. Huyett A, Kahana J, Silver P, Zeng X, Saunders WS: The Kar3p and Kip2p motors function antagonistically at the spindle poles to influence cytoplasmic microtubule numbers. J Cell Sci 1998, 111:295-301.
57. Severin FF, Sorger PK, Hyman AA: Kinetochores distinguish GTP from GDP forms of the microtubule lattice. Nature 1997, 388:888-891.
58. Hoyt MA, Totis L, Roberts BT: S. cerevisiae genes required for cell cycle arrest in response to loss of microtubule function. Cell 1991, 66:507-517.
59. Li R, Murray AW: Feedback control of mitosis in budding yeast. Cell 1991, 66:519-531.
60. Nicklas RB, Ward SC, Gorbsky GJ: Kinetochore chemistry is sensitive to tension and may link mitotic forces to a cell cycle checkpoint. J Cell Biol 1995, 130:929-939.
61. Rieder CL, Schultz A, Cole R, Sluder G: Anaphase onset in vertebrate somatic cells is controlled by a checkpoint that monitors sister kinetochore attachment to the spindle. J Cell Biol 1994, 127:1301-1310.
62. Rieder CL, Cole RW, Khodjakov A, Sluder G: The checkpoint delaying anaphase in response to chromosome monoorientation is mediated by an inhibitory signal produced by unattached kinetochores. J Cell Biol 1995, 130:941-948.
63. Cahill DP, Lengauer C, Yu J, Riggins GJ, Willson JK, Markowitz SD, Kinzler KW, Vogelstein B: Mutations of mitotic checkpoint genes in human cancers. Nature 1998, 392:300-303. Cells exhibiting microsatellite instability (MIN) or chromosomal instability (CIN) were tested for the ability to arrest the cell cycle in the presence of destabilized MTs. Although the MIN lines exhibit normal M phase checkpoint arrest, the CIN lines progress through mitosis without an active checkpoint. The genes for hBUB1 and hBUBR1 were cloned using homology to scBUB1. The coding sequence for hBUB1 is mutated in two out of 19 CIN lines. Heterozygous hBUB1 mutations are observed in the tumors from which the cell lines were derived and not in tumor-free tissue. Expression of mutant hBUB1 results in premature exit from mitosis in nocodazole-treated MIN cells. This work demonstrates an association between aneuploidy in cancerous cells and a mutated checkpoint gene.
64. Farr KA, Hoyt MA: Bub1p kinase activates the Saccharomyces cerevisiae spindle assembly checkpoint. Mol Cell Biol 1998, 18:2738-2747.
65. Bernard P, Hardwick K, Javerzat JP: Fission yeast bub1 is a mitotic centromere protein essential for the spindle checkpoint and the preservation of correct ploidy through mitosis. J Cell Biol 1998, 143:1775-1787. spBUB1 was identified in a Swi6p-deficient screen for lethal mutations. Δbub1 cells are unable to arrest the M→A transition in the presence of destabilized MTs. Segregation of minichromosomes and normal chromosomes were also significantly disrupted in Δbub1 mutant cells in the absence of destabilized MTs. HA epitope tagging was utilized to monitor localization of spBUB1 through the cell cycle. spBUB1 is localized to unattached kinetochores early in the cell cycle and persists in a ΔSwi6 mutant background which presumably contains defective kinetochores.
66. Taylor SS, McKeon F: Kinetochore localization of murine Bub1 is required for normal mitotic timing and checkpoint response to spindle damage. Cell 1997, 89:727-735. mBub1 was cloned by EST homology to scBUB1. Anti-mBUB1 antibodies localize to kinetochores at prophase and prometaphase. At metaphase, only lagging kinetochores stain strongly for mBUB1. Transient transfections with different regions of mBub1 demonstrated that only the amino-terminal domain localized to the kinetochore and expression of this domain interfered with arrest when spindles were disrupted. mBUB1 is a kinase that gets localized to kinetochores before spindle attachment. It subsequently leaves kinetochores that become attached to MTs and remains if MTs do not attach. Therefore mBUB1 may be a signalling protein that enables the segregation apparatus to detect unattached kinetochores.
67. Taylor SS, Ha E, McKeon F: The human homologue of Bub3 is required for kinetochore localization of Bub1 and a Mad3/Bub1-related protein kinase. J Cell Biol 1998, 142:1-11. A hBUB3 cDNA was identified by comparing the scBub3 sequence to a human EST database. A hBUB3-GFP fusion localized hBUB3 to the nucleus during interphase, to kinetochores during prophase and prometaphase and demonstrated that it was absent from kinetochores at metaphase and anaphase. Deletion mutants identified a 38 amino acid region within the amino-terminal domain of mBUB1 that interacts with hBUB3 and MTs. A second EST database search using homology to scMAD3 identified a MAD3/BUB1-related cDNA which is identical to hBUBR1. hBUBR1 also interacts with hBUB3 via the amino-terminal homology domain. A four amino acid region in hBUB3 was shown to be required for binding mBUB1 and hBUBR1.
68. Chen RH, Waters JC, Salmon ED, Murray AW: Association of spindle assembly checkpoint component XMAD2 with unattached kinetochores. Science 1996, 274:242-246.
69. Hardwick KG, Murray AW: Mad1p, a phosphoprotein component of the spindle assembly checkpoint in budding yeast. J Cell Biol 1995, 131:709-720.
70. Li Y, Benezra R: Identification of a human mitotic checkpoint gene: hsMAD2. Science 1996, 274:246-248.
71. Gorbsky GJ: Cell cycle checkpoints: Arresting progress in mitosis. Bioessays 1997, 19:193-197.
72. Chan GK, Schaar BT, Yen TJ: Characterization of the kinetochore binding domain of CENP-E reveals interactions with the kinetochore proteins CENP-F and hBUBR1. J Cell Biol 1998, 143:49-63. A series of deletion constructs of CENP-E identified a 350 amino acid kinetochore binding domain close to the carboxyl terminus. A yeast two-hybrid screen identified an interaction between hBUBR1 and the kinetochore-binding domain of CENP-E. Antibodies to hBUBR1 demonstrate cytoplasmic localization at interphase and at some kinetochores from early prophase. hBUBR1 and CENP-E exhibit coincident localization at prometaphase. CENP-E and hBUBR1 staining was more prominent on chromosomes late to congress and on the trailing kinetochore within pairs of kinetochores on the same congressing chromosome. Staining for both proteins is reduced at kinetochores by mid-anaphase when the colocalization stops. hBUBR1 localizes to the kinetochore before CENP-E and the proteins interact. These results provide evidence for a temporal order to kinetochore formation and a link between a motor kinetochore protein (CENP-E) and a cell-cycle checkpoint protein (hBUBR1).
73. Hwang LH, Lau LF, Smith DL, Mistrot CA, Hardwick KG, Hwang ES, Amon A, Murray AW: Budding yeast Cdc20: A target of the spindle checkpoint. Science 1998, 279:1041-1044. A yeast two-hybrid screen demonstrates an interaction between the checkpoint proteins MAD1, MAD2, MAD3 and the cell-cycle regulator Cdc20p. MAD2 and MAD3 interact directly with Cdc20p whereas the MAD1 interaction with Cdc20p is indirect. A series of mutant strain combinations demonstrated that MAD1 and MAD2 interact directly and they bind MAD3 via MAD2. Although MAD2 and MAD3 interact directly with Cdc20p, this interaction is MAD1 dependent. The significance of this work is that it demonstrates the presence of a CPC and provides a link between checkpoint proteins and the cell cycle arrest apparatus.
74. Fang G, Yu H, Kirschner MW: Direct binding of CDC20 protein family members activates the anaphase-promoting complex in mitosis and G1. Mol Cell 1998, 2:163-171.
75. Visintin R, Prinz S, Amon A: CDC20 and CDH1: A family of substrate-specific activators of APC-dependent proteolysis. Science 1997, 278:460-463.
76. Kim SH, Lin DP, Matsumoto S, Kitazono A, Matsumoto T: Fission yeast Slp1: An effector of the Mad2-dependent spindle checkpoint. Science 1998, 279:1045-1047. A yeast two-hybrid system demonstrates that spMAD2 interacts with Slp1 p. A 29 amino acid domain in Slp1p is important for this interaction; mutations within this region abolish the MAD2-Slp1p interaction and prevent the arrest that would normally be induced by overexpression of spMAD2. This work establishes a link between a checkpoint protein and the cell cycle arrest apparatus in S. pombe.
77. Fang G, Yu H, Kirschner MW: The checkpoint protein MAD2 and the mitotic regulator CDC20 form a ternary complex with the anaphase-promoting complex to control anaphase initiation. Genes Dev 1998, 12:1871-1883. Recombinant hMAD2 exists as a monomer and a tetramer. Only tetramers were capable of inducing cell-cycle arrest after injection into Xenopus embryos. In vitro, the monomeric form of hMAD2 associates with hCDC20 but does not appear to associate with the APC and has no effect on APC activity. The tetrameric form of hMAD2 associates with hCDC20 and the APC to form a ternary complex in vitro and, when formed, the APC is inactive. This study suggests that it is not simply the association of hMAD2 with hCDC20 that inhibits the APC. They present the novel idea that structural changes with hMAD2 may also be involved in the signal transduction process that is required for checkpoint activation.
78. Kallio M, Weinstein J, Daum JR, Burke DJ, Gorbsky GJ: Mammalian p55CDC mediates association of the spindle checkpoint protein mad2 with the cyclosome/anaphase-promoting complex, and is involved in regulating anaphase onset and late mitotic events. J Cell Biol 1998, 141:1393-1406. Anti-p55CDC antibodies exhibit diffuse staining at interphase. Punctate staining was first observed at kinetochores at late prophase which colocaiized with CREST antibodies. The kinetochore labeling was less intense at anaphase and disappeared at telophase. p55CDC-GFP fusions exhibited more intense labeling at kinetochores that had not aligned at the metaphase plate. Sequential anti-p55CDC immunoprecipitations from mitotic HeLa cells contained MAD2 and the Cdc27p component of the APC. The MAD2-Cdc27p association was p55CDC dependent. p55CDC and MAD2 appear to interact directly. Injection of p55CDC antibodies into PtK1 and HeLa cells caused several unusual mitotic events which suggest that p55CDC has multiple roles or is involved in a process that branches into various functions within mitotic mammalian cells. As well as a nice localization study, this paper demonstrates a connection between a CPC and the APC.
79. Wassmann K, Benezra R: Mad2 transiently associates with an APC/p55Cdc complex during mitosis. Proc Natl Acad Sci USA 1998, 95:11193-11198. Coimmunoprecipitations demonstrate that MAD2 associates with hyperphosphorylated Cdc27p in HeLa cells either in the presence or absence of nocodazole. This association is cell-cycle dependent: they associate at the start of mitosis and disassociate at the end of mitosis. Further coimmunoprecipitation experiments in nocodazole-arrested cells demonstrated that p55CDC associates with MAD2, and that Cdc27p associates with p55CDC. Immunodepletion with anti-MAD2 caused a significant reduction in the association of Cdc27p and p55CDC, thereby confirming the presence of a ternary complex. Some Cdc27p-p55CDC complexes exist without MAD2 associations.
80. Dawson IA, Roth S, Artavanis-Tsakonas S: The Drosophila cell cycle gene fizzy is required for normal degradation of cyclins A and B during mitosis and has homology to the CDC20 gene of Saccharomyces cerevisiae. J Cell Biol 1995, 129:725-737.
81. Lorca T, Castro A, Martinez AM, Vigneron S, Morin N, Sigrist S, Lehner C, Doree M, Labbe JC: Fizzy is required for activation of the APC/cyclosome in Xenopus egg extracts. EMBO J 1998, 17:3565-3575.
82. Sigrist SJ, Lehner CF: Drosophila fizzy-related down-regulates mitotic cyclins and is required for cell proliferation arrest and entry into endocycles. Cell 1997, 90:671-681.
83. Matsumoto T: A fission yeast homolog of CDC20/p55CDC/fizzy is required for recovery from DNA damage and genetically interacts with p34cdc2. Mol Cell Biol 1997, 17:742-750.
84. Nicklas RB, Campbell MS, Ward SC, Gorbsky GJ: Tension-sensitive kinetochore phosphorylation in vitro. J Cell Sci 1998, 111:3189-3196. This paper extends previous in vivo studies by demonstrating that kinetochore phosphorylation is dependent upon tension across insect spermatocyte kinetochores in vitro. Relaxed kinetochores exhibit more intense staining with 3F3/2 than kinetochores with micromanipulated tension forces. A kinetochore kinase from HeLa cells is capable of phosphorylating insect kinetochores, demonstrating conservation of sensing mechanisms. This system will provide another method for investigating tension sensing/checkpoint signalling mechanisms.
85. Chen RH, Shevchenko A, Mann M, Murray AW: Spindle checkpoint protein Xmad1 recruits Xmad2 to unattached kinetochores. J Cell Biol 1998, 143:283-295. When xMAD2 is immunodepleted from extracts, the M→A checkpoint is inactivated. The addition of recombinant xMAD2 to normal levels did not restore the checkpoint, indicating that immunodepletion removed some other factor(s) involved in checkpoint activation. Anti-xMAD2 immunoprecipitations have an 85 kDa protein (p85) strongly associated with xMAD2. Mass spectrometry generated eight peptides that were used to isolate Xenopus cDNA clones and identify an open reading frame. Sequence alignments and protein structure predictions with scMAD1, hMAD1 and spMAD1 illustrated that the p85 protein complexed with xMAD2 is xMAD1. Xenopus extracts fail to achieve mitotic arrest after injection of anti-MAD1 antibodies indicating that xMAD1 is also involved in the checkpoint. Localization studies demonstrate that both xMAD1 and xMAD2 have dynamic localization patterns and localize to kinetochores that have not congressed to the metaphase plate.
86. Gorbsky GJ, Chen RH, Murray AW: Microinjection of antibody to Mad2 protein into mammalian cells in mitosis induces premature anaphase. J Cell Biol 1998, 141:1193-1205. Immunofluorescence using anti-MAD2 antibodies demonstrate dynamic localization of MAD2 during the cell cycle. At interphase MAD2 localization is nuclear and cytoplasmic, at prophase the nuclear staining is less diffuse, and at early prometaphase MAD2 is at spindle poles, kinetochores and exhibits granular staining in the cytoplasm. At late prometaphase, MAD2 localization is weak on aligned kinetochores but still prominent on unaligned kinetochores. MAD2 is absent from all kinetochores when all the chromosomes become aligned at metaphase. Microinjection of anti-MAD2 antibody into Ptk1 cells caused early resolution of sister chromatid cohesion and the cells entered into anaphase prematurely. An identical result was observed in human foreskin keratinocytes demonstrating that the arrest is not specific to inbred lines. Therefore MAD2 appears to be necessary for the cell-cycle checkpoint in mammalian cells even in the absence of MT-destabilizing drugs.
87. Campbell MS, Gorbsky GJ: Microinjection of mitotic cells with the 3F3/2 anti-phosphoepitope antibody delays the onset of anaphase. J Cell Biol 1995, 129:1195-1204.
88. Shapiro PS, Vaisberg E, Hunt AJ, Tolwinski NS, Whalen AM, McIntosh JR, Ahn NG: Activation of the MKK/ERK pathway during somatic cell mitosis: Direct interactions of active ERK with kinetochores and regulation of the mitotic 3F3/2 phosphoantigen. J Cell Biol 1998, 142:1533-1545.
89. Li X, Nicklas RB: Tension-sensitive kinetochore phosphorylation and the chromosome distribution checkpoint in praying mantid spermatocytes. J Cell Sci 1997, 110:537-545.
90. Waters JC, Chen RH, Murray AW, Salmon ED: Localization of Mad2 to kinetochores depends on microtubule attachment, not tension. J Cell Biol 1998, 141:1181-1191. MAD2 is localized to unattached kinetochores up until late metaphase when it is lost from attached kinetochores. The authors used the localization of MAD2 to address whether a facet of the M→A checkpoint is to monitor attachment of MTs to the kinetochore or to detect tension across the kinetochore. 'Double-dot' labeling of Ptk1 kinetochores using CREST serum demonstrated that cells treated with taxol (a MT-stabilizing drug) had less space between kinetochores than cells that were not treated with taxol, demonstrating that there is tension across attached kinetochores. Taxol-treated cells with kinetochores attached to MTs did not bind MAD2 even though there was reduced tension. This demonstrates that MAD2 localization in these cells is determined by MT attachment to kinetochores and not tension at the kinetochore. Kinetochores with the fewest MTs had the most MAD2.
91. Biggins S, Severin FF, Bhalla N, Sassoon I, Hyman AA, Murray AW: The conserved protein kinase łpl1 regulates microtubule binding to kinetochores in budding yeast. Genes Dev 1999, 13:532-544.
92. Elledge SJ: Mitotic arrest: Mad2 prevents sleepy from waking up the APC. Science 1998, 279:999-1000.
93. Jorgensen PM, Brundell E, Starborg M, Hoog C: A subunit of the anaphase-promoting complex is a centromere-associated protein in mammalian cells. Mol Cell Biol 1998, 18:468-476. Immunolocalization studies demonstrate that anti-Tsg24 antibodies localization is cell-type dependent. In CHO cells, Tsg24 is absent from kinetochores at interphase and anaphase and colocalize with CREST sera from prophase to early anaphase. Western-blot studies demonstrated that the transient localization of Tsg24 in CHO cells was not caused by transient expression. Contrary to CHO cells, Tsg24 is present at kinetochores throughout mitosis, including late anaphase in two murine cell lines, Swiss-3T3 and L cells. They propose that the transient vs. constitutive localizations could be caused by masking of epitopes in CHO cells. This is a very important paper because it opens the possibility that the APC or components of the APC associate with checkpoint protein complexes at kinetochores.
94. Zachariae W, Shevchenko A, Andrews PD, Ciosk R, Galova M, Stark MJ, Mann M, Nasmyth K: Mass spectrometric analysis of the anaphase-promoting complex from yeast: Identification of a subunit related to cullins. Science 1998, 279:1216-1219.
95. Yanagida M: Fission yeast cut mutations revisited: Control of anaphase. Trends Cell Biol 1998, 8:144-149.
96. Peters JM, King RW, Hoog C, Kirschner MW: Identification of BIME as a subunit of the anaphase-promoting complex. Science 1996, 274:1199-1201.
97. Rieder CL, Khodjakov A, Paliulis LV, Fortier TM, Cole RW, Sluder G: Mitosis in vertebrate somatic cells with two spindles: Implications for the metaphase/anaphase transition checkpoint and cleavage. Proc Natl Acad Sci USA 1997, 94:5107-5112.
98. Ciosk R, Zachariae W, Michaelis C, Shevchenko A, Mann M, Nasmyth K: An ESP1/PDS1 complex regulates loss of sister chromatid cohesion at the metaphase to anaphase transition in yeast. Cell 1998, 93:1067-1076.
99. Kumada K, Nakamura T, Nagao K, Funabiki H, Nakagawa T, Yanagida M: Cut1 is loaded onto the spindle by binding to Cut2 and promotes anaphase spindle movement upon Cut2 proteolysis. Curr Biol 1998, 8:633-641.
100. Mackay AM, Ainsztein AM, Eckley DM, Earnshaw WC: A dominant mutant of inner centromere protein (INCENP), a chromosomal protein, disrupts prometaphase congression and cytokinesis. J Cell Biol 1998, 140:991-1002.
101. Ekwall K, Nimmo ER, Javerzat JP, Borgstrom B, Egel R, Cranston G, Allshire R: Mutations in the fission yeast silencing factors clr4+ and rik1 + disrupt the localisation of the chromo domain protein Swi6p and impair centromere function. J Cell Sci 1996, 109:2637-2648.
102. Torok T, Harvie PD, Buratovich M, Bryant PJ: The product of proliferation disrupter is concentrated at centromeres and required for mitotic chromosome condensation and cell proliferation in Drosophila. Genes Dev 1997, 11:213-225.
103. Kalitsis P, MacDonald AC, Newson AJ, Hudson DF, Choo KH: Gene structure and sequence analysis of mouse centromere proteins A and C. Genomics 1998, 47:108-114.
104. 1/S and forms specialized chromatin required for equal segregation. Cell 1997, 90:131-143.
105. Moore DP, Page AW, Tang TT, Kerrebrock AW, Orr-Weaver TL: The cohesion protein MEI-S332 localizes to condensed meiotic and mitotic centromeres until sister chromatids separate. J Cell Biol 1998, 140:1003-1012.
106. Bickel SE, Wyman DW, Orr-Weaver TL: Mutational analysis of the Drosophila sister-chromatid cohesion protein ORD and its role in the maintenance of centromeric cohesion. Genetics 1997, 146:1319-1331.
107. Maney T, Hunter AW, Wagenbach M, Wordeman L: Mitotic centromere-associated kinesin is important for anaphase chromosome segregation. J Cell Biol 1998, 142:787-801.
108. Cadwell C, Yoon HJ, Zebarjadian Y, Carbon J: The yeast nucleolar protein Cbf5p is involved in rRNA biosynthesis and interacts genetically with the RNA polymerase I transcription factor RRN3. Mol Cell Biol 1997, 17:6175-6183.
109. Starr DA, Williams BC, Li Z, Etemad-Moghadam B, Dawe RK, Goldberg ML: Conservation of the centromere/kinetochore protein ZW10. J Cell Biol 1997, 138:1289-1301.
110. Desai A, Deacon HW, Walczak CE, Mitchison TJ: A method that allows the assembly of kinetochore components onto chromosomes condensed in clarified Xenopus egg extracts. Proc Natl Acad Sci USA 1997, 94:12378-12383.
111. Jin DY, Spencer F, Jeang KT: Human T cell leukemia virus type 1 oncoprotein Tax targets the human mitotic checkpoint protein MAD1. Cell 1998, 93:81-91.
112. Juang YL, Huang J, Peters JM, McLaughlin ME, Tai CY, Pellman D: APC-mediated proteolysis of Ase1 and the morphogenesis of the mitotic spindle. Science 1997, 275:1311-1314.
113. Amon A: Regulation of B-type cyclin proteolysis by Cdc28-associated kinases in budding yeast. EMBO J 1997, 16:2693-2702.
114. Stratmann R, Lehner CF: Separation of sister chromatids in mitosis requires the Drosophila pimples product, a protein degraded after the metaphase/anaphase transition. Cell 1996, 84:25-35.
115. Philp AV, Glover DM: Mutations affecting chromatid separation in Drosophila: The fizzy metaphase arrest persists in pimples fizzy and fizzy three rows double mutants. Exp Cell Res 1997, 230:103-110.
116. Tsuchiya E, Hosotani T, Miyakawa T: A mutation in NPS1/STH1, art essential gene encoding a component of a novel chromatin-remodeling complex RSC, alters the chromatin structure of Saccharomyces cerevisiae centromeres. Nucleic Acids Res 1998, 26:3286-3292.
117. Basrai MA, Kingsbury J, Koshland D, Spencer F, Hieter P: Faithful chromosome transmission requires Spt4p, a putative regulator of chromatin structure in Saccharomyces cerevisiae. Mol Cell Biol 1996, 16:2838-2847.
118. Halverson D, Baum M, Stryker J, Carbon J, Clarke L: A centromere DNA-binding protein from fission yeast affects chromosome segregation and has homology to human CENP-B. J Cell Biol 1997, 136:487-500.
119. Lee JK, Huberman JA, Hurwitz J: Purification and characterization of a CENP-B homologue protein that binds to the centromeric K-type repeat DNA of Schizosaccharomyces pombe. Proc Natl Acad Sci USA 1997, 94:8427-8432.
120. Perrin L, Demakova O, Fanti L, Kallenbach S, Saingery S, Mal'ceva NI, Pimpinelli S, Zhimulev I, Pradel J: Dynamics of the sub-nuclear distribution of Modulo and the regulation of position-effect variegation by nucleolus in Drosophila. J Cell Sci 1998, 111:2753-2761.
121. Logarinho E, Sunkel CE: The Drosophila POLO kinase localises to multiple compartments of the mitotic apparatus and is required for the phosphorylation of MPM2 reactive epitopes. J Cell Sci 1998, 111:2897-2909.
122. Brown KE, Guest SS, Smale ST, Hahm K, Merkenschlager M, Fisher AG: Association of transcriptionally silent genes with Ikaros complexes at centromeric heterochromatin. Cell 1997, 91:845-854.
123. Hahm K, Cobb BS, McCarty AS, Brown KE, Klug CA, Lee R, Akashi K, Weissman IL, Fisher AG, Smale ST: Helios, a T cell-restricted Ikaros family member that quantitatively associates with Ikaros at centromeric heterochromatin. Genes Dev 1998, 12:782-796.
124. 2 and is rapidly degraded after mitosis. J Cell Biol 1995, 130:507-518.
125. Saurin AJ, Shiels C, Williamson J, Satijn DP, Otte AP, Sheer D, Freemont PS: The human polycomb group complex associates with pericentromeric heterochromatin to form a novel nuclear domain. J Cell Biol 1998, 142:887-898.
126. Basu J, Logarinho E, Herrmann S, Bousbaa H, Li Z, Chan GKT, Yen TJ, Sunkel, CE, Goldberg ML: Localization of the Drosophila checkpoint control protein bub3 to the kinetochore requires bub1 but not Zw10 or Rod. Chromosoma 1998, 107:376-385.