Abstract
The composition of the mammalian kinetochore/centromere was studied by indirect immunofluorescence and immunoblotting protocols using serum from a patient with the CREST variant of scleroderma. The results of these studies suggest that a protein with a molecular weight of 50 kDa is localized at the surface of the primary constrictions (the kinetochore region) of both human and Indian muntjac chromosomes. In addition, we were able to verify the presence of a 19.5 kDa antigen (CENP-A), previously detected in human centromeres, within the kinetochore region of the Indian muntjac. These data suggest that the composition of the kinetochore region of the primary constriction is complex and that there is conservation in composition within the mammals. These features may reflect the important role of this unique chromosomal domain in the maintenance of ploidy.
Similar content being viewed by others
References
Ayer LM, Fritzler MJ (1984) Anticentromere antibodies bind to trout testis histone 1 and a low molecular weight protein from rabbit thymus. Mol Immunol 21:761–770
Berke B, Griffiths G, Louvard D, Roggio H, Warren G (1982) A monoclonal antibody against a 135-K golgi membrane protein. EMBO J 1:1621–1628
Blake MS, Johnston KH, Russell-Jones GJ, Gotschlich EC (1984) A rapid-sensitive method for detection of alkaline phosphotase conjugated autoantibody on western blots. Anal Biochem 136:175–179
Brinkley BR, Stubblefield E (1970) Ultrastructure and interaction of the kinetochore and centriole in mitosis and meiosis. Adv Cell Biol 1:119–185
Brinkley BR, Valdivia MM, Tousson A, Brenner AJ (1984) Compound kinetochores of the Indian muntjac. Evolution by linear fusion of unit kinetochores. Chromosoma 91:1–11
Commings DE, Okada TA (1971) Fine structure of the kinetochore in the Indian muntjac. Exp Cell Res 67:97–110
Cox VJ, Schenk EA, Olmsted JB (1983) Human anticentromere antibodies: distribution, characterization of antigens and effect on microtubule organization. Cell 35:331–338
Earnshaw WC, Rothfield N (1985) The identification of a family of human centromere proteins using autoimmune sera from patients with scleroderma. Chromosoma 91:313–319
Earnshaw WC, Bordwell C, Marino C, Rothfield N (1985) Three human chromosomal autoantigens are recognized by sera from patients with anti-centromere antibodies. J Clin Invest 77:426–429
Fritzler MJ, Kinsella TD, Garbutt E (1980) The CREST syndrome: a distinct serological entity with anticentromere antibodies. Am J Med 69:520–523
Guldner HH, Lakomek HL, Bautz FA (1984) Human anti-centromere sera recognize a 19.5 kD non-histone chromosomal protein from HeLa cells. Clin Exp Immunol 58:13–19
Hawkes R (1982) Identification of conconavalin A-binding proteins after sodium dodecyl sulfate-gel electrophoresis and protein blotting. Anal Biochem 123:143–146
Johnson DA, Gautsch JW, Spotsman JR, Elder JH (1984) Improved technique utilizing nonfat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose. Gene Anal Tech 1:3–8
Jokelainen PT (1967) The ultrastructure and spatial organization of the metaphase kinetochore in mitotic rat cells. J Ultrastruct Res 19:19–40
Krishan A, Buck RC (1965) Structure of the mitotic spindle of L strain fibroblasts. J Cell Biol 24:453–444
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the bacteriophage head T4. Nature 227:680–685
Lewis CD, Laemmli UK (1982) Higher order metaphase chromosome structure: evidence for metalloprotein interactions. Cell 29:171–181
McNeilage JL, Whittingham S, McHugh N, Barnett AJ (1986) A highly conserved 72,000 dalton centromeric antigen reactive with autoantibodies from patients with progressive systemic sclerosis. J Immunol 86:2541–2547
Mitchison T, Evans L, Schulze E, Kirschner M (1986) Sites of microtubule assembly and disassembly in the mitotic spindle. Cell 45:515–527
Moroi Y, Peebles C, Fritzler ML, Steigerwald J, Tan EM (1980) Autoantibodies to centromere (kinetochore) in scleroderma sera. Proc Natl Acad Sci USA 77:1627–1631
Nishikai M, Okano Y, Yamashita H, Watanabe M (1984) Characterization of centromere (kinetochore) antigen reactive with sera of patients with a scleroderma variant (CREST syndrome). Ann Rheum Dis 43:819–824
Olmsted JB (1981) Affinity purification of antibodies from diazotized paper blots of heterogeneous protein samples. J Biol Chem 256:1195–57
Palmer DK, O'Day K, Wener MH, Andrews BS, Margolis RL (1987) A 17-kD centromere protein (CENP-A) copurifies with nucleosome core particles and with histones. J Cell Biol 104:805–815
Rattner JB (1986) Organization within the mammalian kinetochore. Chromosoma 93:515–520
Rattner JB (1987) The structure of the kinetochore: A scanning electron microscope study. Chromosoma 95:175–181
Roos UP (1973) Light and electron microscopy of rat kangaroo cells in mitosis. I. Formation and breakdown of the mitotic apparatus. Chromosoma 41:195–220
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
Towbin HS, Staehelin S, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedures and some applications. Proc Natl Acad Sci USA 76:4350–4355
Valdivia MM, Brinkley BR (1985) Fractionation and initial characterization of the kinetochore from mammalian metaphase chromosomes. J Cell Biol 101:1124–29
Wurster DH, Benirschke K (1970) Muntiacus muntjac. A deer with a low diploid chromosome number. Science 186:1364–1366
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kingwell, B., Rattner, J.B. Mammalian kinetochore/centromere composition: a 50 kDa antigen is present in the mammalian kinetochore/centromere. Chromosoma 95, 403–407 (1987). https://doi.org/10.1007/BF00333991
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00333991