Abstract
The organization within the mammalian kinetochore was examined using whole-mount electron microscopic techniques on chromosomes digested with restriction enzymes or micrococcal nuclease. These preparations revealed that a portion of the kinetochore is highly resistant to nuclease digestion and can be visualized as a discrete structure. The relationship of this structure to the remainder of the chromosome suggests that it represents the outer kinetochore plate. The plate is composed of a series of fibrillar loops that are arranged in a parallel array along the plane of the plate. These fibers are 25–30 nm in diameter. The morphology, particulate substructure, and ultimate susceptibility to nuclease digestion suggest that these fibers contain DNA. A model is presented that suggests that the outer plate contains the apexes of chromatin loops that originate within the body of the primary constriction.
Similar content being viewed by others
References
Ayer L, Fritzler M (1984) Anticentromere antibodies bind histones. Mol Immunol 21:761–770
Bloom SK, Amaya E, Carbon J, Clarke L, Hill A, Yeh E (1984) Chromatin conformation in yeast centromeres. J Cell Biol 66:1559–1568
Bloom KS, Fitzgerald-Hayes M, Carbon J (1983) Structural analysis and sequence organization on yeast centromeres. Cold Spring Harbor Symp Quant Biol 47:1175–1185
Brenner S, Pepper D, Berns MW, Tan EM, Brinkley BR (1981) Kinetochore structure, duplication and distribution in mammalian cells. Analysis by human autoantibodies from scleroderma patients. J Cell Biol 91:95–102
Brinkley BR, Stubblefield E (1970) Ultrastructure and interaction of the kinetochore and centriole in mitosis and meiosis. In: Prescott DM, Goldstein L, McConkey E (eds) Advances in cell biology, Vol 1. Appleton Century Croft, New York, pp 119–185
Brinkley BR, Valdivia MM, Tousson A, Brenner AL (1984) Compound kinetochores of the Indian muntjac. Evolution by linear fusion of unit kinetochores. Chromosoma 91:1–11
Clark L, Carbon J (1980) Genomic substitution of centromeres in Saccharomyces cerevisiae. Nature 304:23–38
Commings DE, Okada TA (1971) Fine structure of the kinetochore in the Indian muntjac. Exp Cell Res 67:97–110
Earnshaw WC, Halligan N, Cooke C, Rothfield N (1984) The kinetochore is part of the metaphase chromosome scaffold. J Cell Biol 98:352–357
Fitzgerald-Hayes M, Clarke L, Carbon J (1982) Nucleotide sequences comparisons and functional analysis of yeast centromere DNA. Cell 29:235–244
Jokelaninen PT (1967) The ultrastructure and spatial organization of the metaphase kinetochore in mitotic rat cells. J Ultrastruct Res 19:19–44
Laemmli UK, Cheng SM, Adolph KW, Paulson JR, Brown JA, Baumbach WR (1979) Metaphase chromosome structure: the role of non-histone proteins. Cold Spring Harbor Symp Quant Biol 42:109–118
Lica L, Hamkalo B (1983) Preparation of centromeric heterochromatin by restriction endonuclease digestion of mouse L929 cells. Chromosoma 88:42–49
Maine CT, Surosky RT, Tye BK (1984) Isolation and characterization of centromere from chromosome V (CEN 5) of Saccharomyces cerevisiae. Mol Cell Biol 4:86–91
Matthey R (1973) The chromosome formulae of eutherian mammals. In: Charelli AB, Capanna E (eds) Cytotaxomy and vertebrate evolution. Academic Press, London, pp 531–553
Palmer DK, Margolis RL (1985) Kinetochore components recognized by human autoantibodies are present on mononucleosomes. Mol Cell Biol 5:173–186
Panzeri L, Philipsen P (1982) Centromeric DNA from chromosome VI in Saccharyomes cerevisiae. EMBO J 1:1605–1611
Rattner JB, Branch A, Hamkalo BA (1975) Electron microscopy of whole mount metaphase chromosomes. Chromsoma 66:259–268
Rattner JB, Hamkalo BA (1978) Higher order structure in metaphase chromosomes. I. The 250Å fiber. Chromosoma 69:363–372
Rattner JB, Krystal G, Hamkalo BA (1978) Selective digestion of mouse metaphase chromosomes. Chromosoma 66:259–268
Rattner JB, Lin CC (1985) Radial loops and helical coils co-exist in metaphase chromosomes. Cell, in press
Rieder CL (1981) The structure of cold stable kinetochore fiber in metaphase P+K1 cells. Chromosoma 84:145–158
Rieder CL (1982) The formation, structure and composition of the mammalian kinetochore and kinetochore fiber. Int Rev Cytol 79:1–58
Ris H, Witt PL (1981) Structure of the mammalian kinetochore. Chromosoma 82:153–170
Roos UP (1973) Light and electron microscopy of rat kangaroo cells on mitosis. I. Formation and breakdown of the mitotic apparatus. Chromosoma 41:195–220
Roos UP (1977) The fibrillar organization of the kinetochore and the kinetochore region of mammalian chromosomes. Cytobiology 16:82–90
Shi L, Ye Y, Daux X (1980) Comparative cytogenetic studies on the red muntjac, Chinese muntjac and their I1 hybrids. Cytogenet Cell Genet 26:22–27
Stinchcomb DT, Mann C, Davis RW (1982) Centromeric DNA from Saccharomyces cerevisiae. J Mol Biol 158:157–179
Thoma F, Koller Th, Klug A (1979) Involvement of histone H1 in the organization of the nucleosome and the salt-dependent substructure of chromatin. J Cell Biol 83:403–427
Wurster DH, Benirschke K (1970) Mantacus muntjac. A deer with a low diploid chromosome number. Science 168:1364–1366
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Rattner, J.B. Organization within the mammalian kinetochore. Chromosoma 93, 515–520 (1986). https://doi.org/10.1007/BF00386793
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00386793