Chromosome Research

, Volume 10, Issue 4, pp 267–277

Localization of human SMC1 protein at kinetochores

  • Heather C. Gregson
  • Aaron A. Van Hooser
  • Alexander R. BallJr.
  • B. R. Brinkley
  • Kyoko Yokomori


Proper cohesion of sister chromatids is prerequisite for correct segregation of chromosomes during cell division. The cohesin multiprotein complex, conserved in eukaryotes, is required for sister chromatid cohesion. Human cohesin is composed of a stable heterodimer of the structural maintenance of chromosomes (SMC) family proteins, hSMC1 and hSMC3, and non-SMC components, hRAD21 and SA1 (or SA2). In yeast, cohesin associates with chromosomes from late G1 to metaphase and is required for the establishment and maintenance of both chromosome arm and centromeric cohesion. However, in human cells, the majority of cohesin dissociates from chromosomes before mitosis. Although it was recently shown that a small amount of hRAD21 localizes to the centromeres during metaphase, the presence of other cohesin components at the centromere has not been demonstrated in human cells. Here we report the mitosis-specific localization of hSMC1 to the kinetochores. hSMC1 is targeted to the kinetochore region during prophase concomitant with kinetochore assembly and remains through anaphase. Importantly, hSMC1 is targeted only to the active centromere on dicentric chromosomes. These results suggest that hSMC1 is an integral component of the functional kinetochore structure during mitosis.

chromosome human SMC1 kinetochore mitosis 


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  1. Chan GK, Schaar BT, Yen TJ (1998) Characterization of the kinetochore binding domain of CENP-E reveals interactions with the kinetochore proteins CENP-F and hBUBR1. J Cell Biol 143: 49-63.PubMedCrossRefGoogle Scholar
  2. Choo KH (2000) Centromerization. Trends Cell Biol 10: 182-188.PubMedCrossRefGoogle Scholar
  3. Cooke CA, Schaar BT, Yen TJ, Earnshaw WC (1997) Localization of CENP-E in the fibrous corona and outer plate of mammalian kinetochores from prometaphase through anaphase. Chromosoma 106: 446-455.PubMedCrossRefGoogle Scholar
  4. Craig J, Earnshaw WC, Vagnarelli P (1999) Mammalian centromeres: DNA sequence, protein composition, and role in cell cycle progression. Exp Cell Res 246: 249-262.PubMedCrossRefGoogle Scholar
  5. Darwiche N, Freeman LA, Strunnikov A (1999) Characterization of the components of the putative mammalian sister chromatid cohesion complex. Gene 233: 39-47.PubMedCrossRefGoogle Scholar
  6. Earnshaw WC, Sullivan KF, Machlin PS et al. (1987) Molecular cloning of cDNA for CENP-B, the major human centromere autoantigen. J Cell Biol 104: 817-829.PubMedCrossRefGoogle Scholar
  7. Earnshaw, WC, Ratrie, Hd, Stetten, G (1989) Visualization of centromere proteins CENP-B and CENP-C on a stable dicentric chromosome in cytological spreads. Chromosoma 98: 1-12.PubMedCrossRefGoogle Scholar
  8. Gregson HC, Schmiesing JA, Kim J-S, Kobayashi T, Zhou S, Yokomori, K (2001) A potential role for human cohesin in mitotic spindle aster assembly. J Biol Chem 276: 47575-47582.PubMedCrossRefGoogle Scholar
  9. Guacci V, Koshland D, Strunnikov A (1997) A direct link between sister chromatid cohesion and chromosome condensation revealed through the analysis of MCD1 in S. cerevisiae. Cell 91: 47-57.PubMedCrossRefGoogle Scholar
  10. Heck MMS (1997) Condensins, cohesions, and chromosome architecture: how to make and break a mitotic chromosome. Cell 91: 5-8.PubMedCrossRefGoogle Scholar
  11. Hirano T (1999) SMC-mediated chromosome mechanics: a conserved scheme from bacteria to vertebrates? Genes Dev 13: 11-19.PubMedGoogle Scholar
  12. Hoque MT, Ishikawa K (2001) Human chromatid cohesin component hRad21 is phosphorylated inMphase and associated with metaphase centromeres. J Biol Chem 276: 5059-5067.PubMedCrossRefGoogle Scholar
  13. Jessberger R, Frei C, Gasser SM (1998) Chromosome dynamics: The SMC protein family. Curr Opin Genes Dev 8: 254-259.CrossRefGoogle Scholar
  14. Liao H, Winkfein RJ, Mack G, Rattner JB, Yen TJ (1995) CENP-F is a protein of the nuclear matrix that assembles onto kinetochores at late G2 and is rapidly degraded after mitosis. J Cell Biol 130: 507-518.PubMedCrossRefGoogle Scholar
  15. Losada A, Hirano T (2000) Biology in pictures. New light on sticky sisters. Curr Biol 10: R615.PubMedCrossRefGoogle Scholar
  16. Losada A, Hirano M, Hirano T (1998) Identification of Xenopus SMC protein complexes required for sister chromatid cohesion. Genes Dev 12: 1986-1997.PubMedGoogle Scholar
  17. Losada A, Yokochi T, Kobayashi R, Hirano T (2000) Identi-fication and characterization of SA/Scc3p subunits in the Xenopus and human cohesin complexes. J Cell Biol 150: 405-416.PubMedCrossRefGoogle Scholar
  18. Megee PC, Koshland D (1999) A functional assay for centromere-associated sister chromatid cohesion. Science 285: 254-257.PubMedCrossRefGoogle Scholar
  19. Megee PC, Mistrot C, Guacci V, Koshland D (1999) The centromeric sister chromatid cohesion site directs Mcd1p binding to adjacent sequences. Mol Cell 4: 445-450.PubMedCrossRefGoogle Scholar
  20. Melby TE, Ciampaglio CN, Briscoe G, Erickson HP (1998) The symmetrical structure of structural maintenance of chromosomes (SMC) and MukB proteins: Long, antiparallel coiled coils, folded at a flexible hinge. J Cell Biol 142: 1595-1604.PubMedCrossRefGoogle Scholar
  21. Merry DE, Pathak S, Hsu TC, Brinkley BR (1985) Antikinetochore antibodies: use as probes for inactive centromeres. Am J Hum Genet 37: 425-430.PubMedGoogle Scholar
  22. Michaelis C, Ciosk R, Nasmyth K (1997) Cohesins: Chromosomal proteins that prevent premature separation of sister chromatids. Cell 91: 35-45.PubMedCrossRefGoogle Scholar
  23. Schmiesing JA, Ball AR, Gregson HC, Alderton J, Zhou S, Yokomori K (1998) Identification of two distinct human SMC protein complexes involved in mitotic chromosome dynamics. Proc Natl Acad Sci USA 95: 12906-12911.PubMedCrossRefGoogle Scholar
  24. Schmiesing JA, Gregson HC, Zhou S, Yokomori K (2000) A human condensin complex containing hCAP-C/hCAP-E and CNAP1, a homolog of Xenopus XCAP-D2, colocalizes with phosphorylated histone H3 during the early stage of mitotic chromosome condensation. Mol Cell Biol 20: 6996-7006.PubMedCrossRefGoogle Scholar
  25. Shelby RD, Vafa O, Sullivan KF (1997) Assembly of CENP-A into centromeric chromatin requires a cooperative array of nucleosomal DNA contact sites. J Cell Biol 136: 501-513.PubMedCrossRefGoogle Scholar
  26. Sumara I, Vorlaufer E, Gieffers C, Peters BH, Peters J-M (2000) Characterization of vertebrate cohesin complexes and their regulation in prophase. J Cell Biol 151: 749-762.PubMedCrossRefGoogle Scholar
  27. Tanaka T, Cosma MP, Wirth K, Nasmyth K (1999) Identifi-cation of cohesin association sites at centromeres and along chromosome arms. Cell 98: 847-858.PubMedCrossRefGoogle Scholar
  28. Tanaka T, Fuchs J, Loidl J, Nasmyth K (2000) Cohesin ensures bipolar attachment of microtubules to sister centromeres and resists their precocious separation. Nat Cell Biol 2: 492-499.PubMedCrossRefGoogle Scholar
  29. Tomonaga T, Nagao K, Kawasaki Y et al. (2000) Characterization of fission yeast cohesin: essential anaphase proteolysis of Rad21 phosphorylated in the S phase. Genes Dev 14: 2757-2770.PubMedCrossRefGoogle Scholar
  30. Toth A, Ciosk R, Uhlmann F, Galova M, Schleiffer A, Nasmyth K (1999) Yeast cohesin complex requires a conserved protein, Eco1p (Ctf7), to establish cohesion between sister chromatids during DNA replication. Genes Dev 13: 320-333.PubMedGoogle Scholar
  31. Uhlmann F, Lottspeich F, Nasmyth K (1999) Sister-chromatid separation at anaphase onset is promoted by cleavage of the cohesin subunit Scc1. Nature 400: 37-42.PubMedCrossRefGoogle Scholar
  32. Uhlmann F, Wernic D, Poupart M-A, Koonin EV, Nasmyth K (2000) Cleavage of cohesin by the CD clan protease separin triggers anaphase in yeast. Cell 103: 375-386.PubMedCrossRefGoogle Scholar
  33. Valdivia MM, Brinkley BR (1985) Fractionation and initial characterization of the kinetochore from mammalian metaphase chromosomes. J Cell Biol 101: 1124-1134.PubMedCrossRefGoogle Scholar
  34. Van Hooser A, Brinkley WR (1999) Methods for in situ localization of proteins and DNA in the centromere-kinetochore complex. Meth Cell Biol 61: 57-80.CrossRefGoogle Scholar
  35. Van Hooser AA, Ouspenski II, Gregson HC et al. (2001) Speci-fication of kinetochore-forming chromatin by the histone H3 variant CENP-A. J Cell Sci 114: 3529-3542.PubMedGoogle Scholar
  36. Vig BK, Rattner, JB (1989) Centromere, kinetochore, and cancer. Crit Rev Oncogen 1: 343-371.Google Scholar
  37. Vig BK, Latour, D, Frankovich, J (1994) Dissociation of minor satellite fromt he centromere in mouse. J Cell Sci 107: 3091-3095.PubMedGoogle Scholar
  38. Waizenegger IC, Hauf S, Meinke A, Peters J-M (2000) Two distinct pathways remove mammalian cohesin from chromosome arms in prophase and from centromeres in anaphase. Cell 103: 399-410.PubMedCrossRefGoogle Scholar
  39. Warburton PE, Cooke CA, Bourassa S et al. (1997) Immunolocalization of CENP-A suggests a distinct nucleosome structure at the inner kinetochore plate of active centromeres. Curr Biol 7: 901-904.PubMedCrossRefGoogle Scholar
  40. Warren W, Steffensen S, Lin E et al. (2000) The Drosophila RAD21 cohesin persists at the centromere region in mitosis. Curr Biol. 10: 1463-6.PubMedCrossRefGoogle Scholar
  41. Yen TJ, Compton DA, Wise D et al. (1991) CENP-E, a novel human centromere-associated protein required for progression fromm etaphase to anaphase. EMBO J 10: 1245-1254.PubMedGoogle Scholar
  42. Zheng L, Chen Y, Lee W-H (1999) Hec1p, an evolutionarily conserved coiled-coil protein, modulates chromosome segregation through interaction with SMC proteins. Mol Biol Cell 19: 5417-5428.Google Scholar

Copyright information

© Kluwer Academic Publishers 2002

Authors and Affiliations

  • Heather C. Gregson
    • 1
  • Aaron A. Van Hooser
    • 2
  • Alexander R. BallJr.
    • 1
  • B. R. Brinkley
    • 2
  • Kyoko Yokomori
    • 1
  1. 1.Department of Biological ChemistryUniversity of California, Irvine, College of MedicineIrvineUSA
  2. 2.Department of Molecular and Cellular BiologyBaylor College of MedicineHoustonUSA
  3. 3.Department of Molecular & Cell BiologyUniversity of California, BerkeleyBerkeleyUSA

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