Abstract
A critical aspect of mitosis is the interaction of the kinetochore with spindle microtubules. Fission yeast Mal3 is a member of the EB1 family of microtubule plus-end binding proteins, which have been implicated in this process. However, the Mal3 interaction partner at the kinetochore had not been identified. Here, we show that the mal3 mutant phenotype can be suppressed by the presence of extra Spc7, an essential kinetochore protein associated with the central centromere region. Mal3 and Spc7 interact physically as both proteins can be coimmunoprecipitated. Overexpression of a Spc7 variant severely compromises kinetochore–microtubule interaction, indicating that the Spc7 protein plays a role in this process. Spc7 function seems to be conserved because, Spc105, a Saccharomyces cerevisiae homolog of Spc7, identified by mass spectrometry as a component of the conserved Ndc80 complex, can rescue mal3 mutant strains.
Bibliography
Kerres, A., Vietmeier-Decker, C., Ortiz, J., Karig, I., Beuter, C., Hegemann, J., Lechner, J., & Fleig, U. (2004). The Fission Yeast Kinetochore Component Spc7 Associates with the EB1 Family Member Mal3 and Is Required for KinetochoreâSpindle Association. Molecular Biology of the Cell, 15(12), 5255â5267.
References
72
Referenced
39
-
Akhmanova, A.et al. (2001). Clasps are CLIP-115 and -170 associating proteins involved in the regional regulation of microtubule dynamics in motile fibroblasts.Cell104, 923–935.
(
10.1016/S0092-8674(01)00288-4
) -
Arnaoutov, A., and Dasso, M. (2003). The Ran GTPase regulates kinetochore function.Dev. Cell5, 99–111.
(
10.1016/S1534-5807(03)00194-1
) -
Bamba, C., Bobinnec, Y., Fukuda, M., and Nishida, E. (2002). The GTPase Ran regulates chromosome positioning and nuclear envelope assembly in vivo.Curr. Biol.12, 503–507.
(
10.1016/S0960-9822(02)00741-8
) -
Barbet, N., Muriel, W.J., and Carr, A.M. (1992). Versatile shuttle vectors and genomic libraries for use withSchizosaccharomyces pombe.Gene114, 59–66.
(
10.1016/0378-1119(92)90707-V
) -
Beinhauer, J.D., Hagan, I.M., Hegemann, J.H., and Fleig, U. (1997). Mal3, the fission yeast homologue of the human APC-interacting protein EB-1 is required for microtubule integrity and the maintenance of cell form.J. Cell Biol.139, 717–728.
(
10.1083/jcb.139.3.717
) -
Bernard, P., and Allshire, R. (2002). Centromeres become unstuck without heterochromatin.Trends Cell Biol.12, 419–424.
(
10.1016/S0962-8924(02)02344-9
) -
Bharadwaj, R., Qi, W., and Yu, H. (2004). Identification of two novel components of the human Ndc80 kinetochore complex.J. Biol. Chem.279, 13076–13085.
(
10.1074/jbc.M310224200
) -
Bridge, A.J., Morphew, M., Bartlett, R., and Hagan, I.M. (1998). The fission yeast SPB component Cut12 links bipolar spindle formation to mitotic control.Genes Dev.12, 927–942.
(
10.1101/gad.12.7.927
) -
Bu, W., and Su, L.K. (2001). Regulation of microtubule assembly by human EB1 family proteins.Oncogene20, 3185–3192.
(
10.1038/sj.onc.1204429
) -
Busch, K.E., and Brunner, D. (2004). The microtubule plus end-tracking proteins mal3p and tip1p cooperate for cell-end targeting of interphase microtubules.Curr. Biol.14, 548–559.
(
10.1016/j.cub.2004.03.029
) -
Cheeseman, I.M., Drubin, D.G., and Barnes, G. (2002). Simple centromere, complex kinetochore: linking spindle microtubules and centromeric DNA in budding yeast.J. Cell Biol.157, 199–203.
(
10.1083/jcb.200201052
) -
Clarke, L. (1998). Centromeres: proteins, protein complexes, and repeated domains at centromeres of simple eukaryotes.Curr. Opin. Genet. Dev.8, 212–218.
(
10.1016/S0959-437X(98)80143-3
) -
Cleveland, D.W., Mao, Y., and Sullivan, K.F. (2003). Centromeres and kinetochores: from epigenetics to mitotic checkpoint signaling.Cell112, 407–421.
(
10.1016/S0092-8674(03)00115-6
) -
De Wulf, P., McAinsh, A.D., and Sorger, P.K. (2003). Hierarchical assembly of the budding yeast kinetochore from multiple subcomplexes.Genes Dev.17, 2902–2921.
(
10.1101/gad.1144403
) -
DeLuca, J.G., Moree, B., Hickey, J.M., Kilmartin, J.V., and Salmon, E.D. (2002). hNuf2 inhibition blocks stable kinetochore-microtubule attachment and induces mitotic cell death in HeLa cells.J. Cell Biol.159, 549–555.
(
10.1083/jcb.200208159
) -
Dujardin, D., Wacker, U.I., Moreau, A., Schroer, T.A., Rickard, J.E., and De Mey, J.R. (1998). Evidence for a role of CLIP-170 in the establishment of metaphase chromosome alignment.J. Cell Biol.141, 849–862.
(
10.1083/jcb.141.4.849
) -
Ekwall, K., Javerzat, J.P., Lorentz, A., Schmidt, H., Cranston, G., and Allshire, R. (1995). The chromodomain protein Swi6, a key component at fission yeast centromeres.Science269, 1429–1431.
(
10.1126/science.7660126
) -
Fleig, U., Salus, S.S., Karig, I., and Sazer, S. (2000). The fission yeast Ran GTPase is required for microtubule integrity.J. Cell Biol.151, 1101–1112.
(
10.1083/jcb.151.5.1101
) -
Fleig, U., Sen-Gupta, M., and Hegemann, J.H. (1996). Fission yeast mal2+ is required for chromosome segregation.Mol. Cell. Biol.16, 6169–6177.
(
10.1128/MCB.16.11.6169
) -
Fodde, R.et al. (2001). Mutations in the APC tumour suppressor gene cause chromosomal instability.Nat. Cell Biol.3, 433–438.
(
10.1038/35070129
) -
Funabiki, H., Hagan, I., Uzawa, S., and Yanagida, M. (1993). Cell cycle-dependent specific positioning and clustering of centromeres and telomeres in fission yeast.J. Cell Biol.121, 961–976.
(
10.1083/jcb.121.5.961
) -
Garcia, M.A., Vardy, L., Koonrugsa, N., and Toda, T. (2001). Fission yeast ch-TOG/XMAP215 homologue Alp14 connects mitotic spindles with the kinetochore and is a component of the Mad2-dependent spindle checkpoint.EMBO J.20, 3389–3401.
(
10.1093/emboj/20.13.3389
) -
Gari, E., Piedrafita, L., Aldea, M., and Herrero, E. (1997). A set of vectors with a tetracycline-regulatable promoter system for modulated gene expression inSaccharomyces cerevisiae.Yeast13, 837–848.
(
10.1002/(SICI)1097-0061(199707)13:9<837::AID-YEA145>3.0.CO;2-T
) -
Giaever, G.et al. (2002). Functional profiling of theSaccharomyces cerevisiaegenome.Nature418, 387–391.
(
10.1038/nature00935
) -
Goshima, G., Saitoh, S., and Yanagida, M. (1999). Proper metaphase spindle length is determined by centromere proteins Mis12 and Mis6 required for faithful chromosome segregation.Genes Dev.13, 1664–1677.
(
10.1101/gad.13.13.1664
) - Gundersen, G.G., and Bretscher, A. (2003). Cell biology. Microtubule asymmetry.Science300, 2040–2041.
-
Hagan, I.M., and Hyams, J.S. (1988). The use of cell division cycle mutants to investigate the control of microtubule distribution in the fission yeastSchizosaccharomyces pombe.J. Cell Sci.89, 343–357.
(
10.1242/jcs.89.3.343
) -
He, X., Jones, M.H., Winey, M., and Sazer, S. (1998). Mph1, a member of the Mps1-like family of dual specificity protein kinases, is required for the spindle checkpoint inS. pombe.J. Cell Sci.111, 1635–1647.
(
10.1242/jcs.111.12.1635
) -
He, X., Rines, D.R., Espelin, C.W., and Sorger, P.K. (2001). Molecular analysis of kinetochore-microtubule attachment in budding yeast.Cell106, 195–206.
(
10.1016/S0092-8674(01)00438-X
) -
Huh, W.K., Falvo, J.V., Gerke, L.C., Carroll, A.S., Howson, R.W., Weissman, J.S., and O'Shea, E.K. (2003). Global analysis of protein localization in budding yeast.Nature425, 686–691.
(
10.1038/nature02026
) -
Janke, C., Ortiz, J., Lechner, J., Shevchenko, A., Magiera, M.M., Schramm, C., and Schiebel, E. (2001). The budding yeast proteins Spc24p and Spc25p interact with Ndc80p and Nuf2p at the kinetochore and are important for kinetochore clustering and checkpoint control.EMBO J.20, 777–791.
(
10.1093/emboj/20.4.777
) -
Jin, Q.W., Pidoux, A.L., Decker, C., Allshire, R.C., and Fleig, U. (2002). The mal2p protein is an essential component of the fission yeast centromere.Mol. Cell. Biol.22, 7168–7183.
(
10.1128/MCB.22.20.7168-7183.2002
) -
Joseph, J., Tan, S.H., Karpova, T.S., McNally, J.G., and Dasso, M. (2002). SUMO-1 targets RanGAP1 to kinetochores and mitotic spindles.J. Cell Biol.156, 595–602.
(
10.1083/jcb.200110109
) -
Juwana, J.P., Henderikx, P., Mischo, A., Wadle, A., Fadle, N., Gerlach, K., Arends, J.W., Hoogenboom, H., Freudschuh, M., and Renner, C. (1999). EB/RP gene family encodes tubulin binding proteins.Int. J. Cancer81, 275–284.
(
10.1002/(SICI)1097-0215(19990412)81:2<275::AID-IJC18>3.0.CO;2-Z
) - Kaiser, C., Michaelis, S., and Mitchell, A. (1994).Methods in Yeast Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.
-
Kaplan, K.B., Burds, A.A., Swedlow, J.R., Bekir, S.S., Sorger, P.K., and Nathke, I.S. (2001). A role for the adenomatous polyposis coli protein in chromosome segregation.Nat. Cell Biol.3, 429–432.
(
10.1038/35070123
) -
Kirschner, M., and Mitchison, T. (1986). Beyond self-assembly: from microtubules to morphogenesis.Cell45, 329–342.
(
10.1016/0092-8674(86)90318-1
) 10.1091/mbc.12.9.2767
10.1091/mbc.e02-03-0155
-
Lin, H., de Carvalho, P., Kho, D., Tai, C.Y., Pierre, P., Fink, G.R., and Pellman, D. (2001). Polyploids require Bik1 for kinetochore-microtubule attachment.J. Cell Biol.155, 1173–1184.
(
10.1083/jcb.200108119
) -
Maiato, H., Fairley, E.A., Rieder, C.L., Swedlow, J.R., Sunkel, C.E., and Earnshaw, W.C. (2003). Human CLASP1 is an outer kinetochore component that regulates spindle microtubule dynamics.Cell113, 891–904.
(
10.1016/S0092-8674(03)00465-3
) -
McCleland, M.L., Kallio, M.J., Barrett-Wilt, G.A., Kestner, C.A., Shabanowitz, J., Hunt, D.F., Gorbsky, G.J., and Stukenberg, P.T. (2004). The vertebrate Ndc80 complex contains Spc24 and Spc25 homologs, which are required to establish and maintain kinetochore-microtubule attachment.Curr. Biol.14, 131–137.
(
10.1016/j.cub.2003.12.058
) -
McIntosh, J.R., Grishchuk, E.L., and West, R.R. (2002). Chromosome-microtubule interactions during mitosis.Annu. Rev. Cell Dev. Biol.18, 193–219.
(
10.1146/annurev.cellbio.18.032002.132412
) -
Mimori-Kiyosue, Y., and Tsukita, S. (2003). “Search-and-capture” of microtubules through plus-end-binding proteins (+TIPs).J. Biochem.134, 321–326.
(
10.1093/jb/mvg148
) -
Moreno, M.B., Duran, A., and Ribas, J.C. (2000). A family of multifunctional thiamine-repressible expression vectors for fission yeast.Yeast16, 861–872.
(
10.1002/1097-0061(20000630)16:9<861::AID-YEA577>3.0.CO;2-9
) -
Moreno, S., Klar, A., and Nurse, P. (1991). Molecular genetic analysis of fission yeastSchizosaccharomyces pombe.Methods Enzymol.194, 795–823.
(
10.1016/0076-6879(91)94059-L
) -
Nabeshima, K., Kurooka, H., Takeuchi, M., Kinoshita, K., Nakaseko, Y., and Yanagida, M. (1995). p93dis1, which is required for sister chromatid separation, is a novel microtubule and spindle pole body-associating protein phosphorylated at the Cdc2 target sites.Genes Dev.9, 1572–1585.
(
10.1101/gad.9.13.1572
) -
Nakamura, M., Zhou, X.Z., and Lu, K.P. (2001). Critical role for the EB1 and APC interaction in the regulation of microtubule polymerization.Curr. Biol.11, 1062–1067.
(
10.1016/S0960-9822(01)00297-4
) -
Nakaseko, Y., Goshima, G., Morishita, J., and Yanagida, M. (2001). M phase-specific kinetochore proteins in fission yeast. Microtubule-associating Dis1 and Mtc1 display rapid separation and segregation during anaphase.Curr. Biol.11, 537–549.
(
10.1016/S0960-9822(01)00155-5
) 10.1091/mbc.e03-06-0419
-
Niwa, O., Matsumoto, T., and Yanagida, M. (1986). Construction of a minichromosome by deletion and its mitotic and meiotic behaviour in fission yeast.Mol. Gen. Genet.203, 397–405.
(
10.1007/BF00422063
) -
Ohkura, H., Adachi, Y., Kinoshita, N., Niwa, O., Toda, T., and Yanagida, M. (1988). Cold-sensitive and caffeine-supersensitive mutants of theSchizosaccharomyces pombedis genes implicated in sister chromatid separation during mitosis.EMBO J.7, 1465–1473.
(
10.1002/j.1460-2075.1988.tb02964.x
) -
Partridge, J.F., Borgstrom, B., and Allshire, R.C. (2000). Distinct protein interaction domains and protein spreading in a complex centromere.Genes Dev.14, 783–791.
(
10.1101/gad.14.7.783
) -
Pidoux, A.L., and Allshire, R.C. (2000). Centromeres: getting a grip of chromosomes.Curr. Opin. Cell Biol.12, 308–319.
(
10.1016/S0955-0674(00)00094-6
) -
Pidoux, A.L., Richardson, W., and Allshire, R.C. (2003). Sim4, a novel fission yeast kinetochore protein required for centromeric silencing and chromosome segregation.J. Cell Biol.161, 295–307.
(
10.1083/jcb.200212110
) 10.1091/mbc.e02-01-0054
-
Rogers, S.L., Rogers, G.C., Sharp, D.J., and Vale, R.D. (2002). Drosophila EB1 is important for proper assembly, dynamics, and positioning of the mitotic spindle.J. Cell Biol.158, 873–884.
(
10.1083/jcb.200202032
) -
Saitoh, S., Takahashi, K., and Yanagida, M. (1997). Mis6, a fission yeast inner centromere protein, acts during G1/S and forms specialized chromatin required for equal segregation.Cell90, 131–143.
(
10.1016/S0092-8674(00)80320-7
) -
Salus, S.S., Demeter, J., and Sazer, S. (2002). The Ran GTPase system in fission yeast affects microtubules and cytokinesis in cells that are competent for nucleocytoplasmic protein transport.Mol. Cell. Biol.22, 8491–8505.
(
10.1128/MCB.22.24.8491-8505.2002
) -
Schuyler, S.C., and Pellman, D. (2001). Microtubule “plus-end-tracking proteins”: the end is just the beginning.Cell105, 421–424.
(
10.1016/S0092-8674(01)00364-6
) 10.1091/mbc.8.12.2677
-
Shevchenko, A., Wilm, M., Vorm, O., and Mann, M. (1996). Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels.Anal. Chem.68, 850–858.
(
10.1021/ac950914h
) - Su, L.K., Burrell, M., Hill, D.E., Gyuris, J., Brent, R., Wiltshire, R., Trent, J., Vogelstein, B., and Kinzler, K.W. (1995). APC binds to the novel protein EB1.Cancer Res.55, 2972–2977.
-
Tirnauer, J.S., and Bierer, B.E. (2000). EB1 Proteins Regulate Microtubule Dynamics, Cell Polarity, and Chromosome Stability.J. Cell Biol.149, 761–766.
(
10.1083/jcb.149.4.761
) 10.1091/mbc.e02-04-0236
10.1091/mbc.e02-04-0210
-
Tirnauer, J.S., O'Toole, E., Berrueta, L., Bierer, B.E., and Pellman, D. (1999). Yeast Bim1p promotes the G1-specific dynamics of microtubules.J. Cell Biol.145, 993–1007.
(
10.1083/jcb.145.5.993
) -
Umesono, K., Toda, T., Hayashi, S., and Yanagida, M. (1983). Cell division cycle genes nda2 and nda3 of the fission yeastSchizosaccharomyces pombecontrol microtubular organization and sensitivity to anti-mitotic benzimidazole compounds.J. Mol. Biol.168, 271–284.
(
10.1016/S0022-2836(83)80018-7
) -
Wigge, P.A., Jensen, O.N., Holmes, S., Soues, S., Mann, M., and Kilmartin, J.V. (1998). Analysis of theSaccharomycesspindle pole by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry.J. Cell Biol.141, 967–977.
(
10.1083/jcb.141.4.967
) -
Wigge, P.A., and Kilmartin, J.V. (2001). The Ndc80p complex fromSaccharomyces cerevisiaecontains conserved centromere component and has a function in chromosome segregation.J. Cell Biol.152, 349–360.
(
10.1083/jcb.152.2.349
) -
Woods, A., Sherwin, T., Sasse, R., MacRae, T.H., Baines, A.J., and Gull, K. (1989). Definition of individual components within the cytoskeleton ofTrypanosoma bruceiby a library of monoclonal antibodies.J. Cell Sci.93, 491–500.
(
10.1242/jcs.93.3.491
) -
Yamada, H., Kumada, K., and Yanagida, M. (1997). Distinct subunit functions and cell cycle regulated phosphorylation of 20S APC/cyclosome required for anaphase in fission yeast.J. Cell Sci.110, 1793–1804.
(
10.1242/jcs.110.15.1793
)
Dates
Type | When |
---|---|
Created | 20 years, 11 months ago (Sept. 15, 2004, 8:28 p.m.) |
Deposited | 2 years, 4 months ago (April 29, 2023, 5:41 a.m.) |
Indexed | 4 months, 2 weeks ago (April 17, 2025, 7:27 a.m.) |
Issued | 20 years, 9 months ago (Dec. 1, 2004) |
Published | 20 years, 9 months ago (Dec. 1, 2004) |
Published Print | 20 years, 9 months ago (Dec. 1, 2004) |
@article{Kerres_2004, title={The Fission Yeast Kinetochore Component Spc7 Associates with the EB1 Family Member Mal3 and Is Required for Kinetochore–Spindle Association}, volume={15}, ISSN={1939-4586}, url={http://dx.doi.org/10.1091/mbc.e04-06-0443}, DOI={10.1091/mbc.e04-06-0443}, number={12}, journal={Molecular Biology of the Cell}, publisher={American Society for Cell Biology (ASCB)}, author={Kerres, Anne and Vietmeier-Decker, Corina and Ortiz, Jennifer and Karig, Inga and Beuter, Christoph and Hegemann, Johannes and Lechner, Johannes and Fleig, Ursula}, year={2004}, month=dec, pages={5255–5267} }