Skip to main content
SpringerLink
Log in

Molecular-cytogenetic characterization of a higher plant centromere/kinetochore complex

  • Published:
Theoretical and Applied Genetics Aims and scope Submit manuscript

Abstract

The centromeric region of a telocentric field bean chromosome that resulted from centric fission of the metacentric satellite chromosome was microdissected. The DNA of this region was amplified and biotinylated by degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR)/linker-adapter PCR. After fluorescence in situ hybridization (FISH) the entire chromosome complement of Vicia faba was labelled by these probes except for the nucleolus organizing region (NOR) and the interstitial heterochromatin, the chromosomes of V. sativa and V. narbonensis were only slightly labelled by the same probes. Dense uniform labelling was also observed when a probe amplified from a clearly delimited microdissected centromeric region of a mutant of Tradescantia paludosa was hybridized to T. paludosa chromosomes. Even after six cycles of subtractive hybridization between DNA fragments amplified from centromeric and acentric regions no sequences specifically located at the field bean centromeres were found among the remaining DNA. A mouse antiserum was produced which detected nuclear proteins of 33 kDa and 68 kDa; these were predominantly located at V. faba kinetochores during mitotic metaphase. DNA amplified from the chromatin fraction adsorbed by this serum out of the sonicated total mitotic chromatin also did not cause specific labelling of primary constrictions. From these results we conclude: (1) either centromere-specific DNA sequences are not very conserved among higher plants and are — at least in species with large genomes — intermingled with complex dispersed repetitive sequences that prevent the purification of the former, or (2) (some of) the dispersed repeats themselves specify the primary constrictions by stereophysical parameters rather than by their base sequence.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Alfenito MR, Birchler JA (1993) Molecular characterization of a maize B chromosome centric sequence. Genetics 135:589–597

    Google Scholar 

  • Archidiacono N, Antonacci R, Marzella R, Finelli P, Lonoce A, Rocchi M (1995) Comparative mapping of human alphoid sequences in great apes using fluorescence situ hybridization. Genomics 25:477–484

    Google Scholar 

  • Binarova P, Cihalikova J, Dolezel J (1993) Localization of MPM-2 recognized phosphoproteins and tubulin during cell cycle progression in synchronized Vicia faba root meristem cells. Cell Biol Int 17:847–856

    Google Scholar 

  • Brenner S, Peper D, Berns MW, Tan E, Brinkley BR (1981) Kinetochore structure, duplication, and distribution in mammalian cells: analysis by human autoantibodies from scleroderma patients. J Cell Biol 91:95–102

    Google Scholar 

  • Clarke L, Baum M, Marschall LG, Ngan VK, Steiner NC (1993) Structure and function of Schizosaccharomyces pombe centromeres. Cold Spring Harbor Symp Quant Biol 58:687–695

    Google Scholar 

  • Döbel P, Schubert I, Rieger R (1978) Distribution of heterochromatin in a reconstructed karyotype of Vicia faba as identified by banding- and DNA-late replication patterns. Chromosoma 69:193–209

    Google Scholar 

  • Earnshaw WC (1991) When is a centromere not a kinetochore? J Cell Sci 99:1–4

    Google Scholar 

  • Earnshaw WC, MacKay AM (1994) Role of nonhistone proteins in the chromosomal events of mitosis. FASEB J 8:947–956

    Google Scholar 

  • Earnshaw WC, Rothfield N (1985) Identification of a family of human centromere proteins using autoimmune sera from patients with scleroderma. Chromosoma 91:313–321

    Google Scholar 

  • Fuchs J, Schubert I (1995) Localization of seed protein genes on metaphase chromosomes of Vicia faba via fluorescence situ hybridization. Chromosome Res 3:94–100

    Google Scholar 

  • Fuchs J, Pich U, Meister A, Schubert I (1994) Differentiation of field bean heterochromatin by in situ hybridization with a repeated FokI sequence. Chromosome Res 2:25–28

    Google Scholar 

  • Fuchs J, Houben A, Brandes A, Schubert I (1996) Chromosome ‘painting’ in plants — a feasible technique? Chromosoma 104:315–320

    Google Scholar 

  • Haaf T, Warburton PE, Willard HF (1992) Integration of human α-satellite DNA into simian chromosomes: centromere protein binding and disruption of normal chromosome segregation. Cell 70:681–696

    Google Scholar 

  • Hadlaczky G, Went M, Ringertz NR (1986) Direct evidence for the non-random localization of mammalian chromosomes in the interphase nucleus. Exp Cell Res 167:1–15

    Google Scholar 

  • Hadlaczky G, Praznovszky T, Cserpán I, Keresö J, Péterfy M, Kelemen I, Atalay E, Szeles A, Szelei J, Tubak V, Burg K (1991) Centromere formation in mouse cells cotransformed with human DNA and a dominant marker gene. Proc Natl Acad Sci USA 88:8106–8110

    Google Scholar 

  • Harrison GE, Heslop-Harrison JS (1995) Centromeric repetitive DNA sequences in the genus Brassica. Theor Appl Genet 90:157–165

    Google Scholar 

  • Heus JJ, Bloom KS, Zonneveld BJM, Steensma HY, Van den Berg JA (1993) Chromatin structures of Kluyveromyces lactis centromeres in K. lactis and Saccharomyces cerevisiae. Chromosoma 102:660–667

    Google Scholar 

  • Houben A, Guttenbach M, Kreß W, Pich U, Schubert I, Schmid M (1995) Immunostaining and interphase arrangement of field bean kinetochores. Chromosome Res 3:27–31

    Google Scholar 

  • Houben A, Franke J, Leclerc N, Ahne R (1996) A computer assisted system combining image analysis and chromosome dissection. Microsc Res Technol (in press)

  • Jiang J, Gill BS (1994) Non-isotopic in situ hybridization and plant genome mapping: the first 10 years. Genome 37:717–725

    Google Scholar 

  • Johnson DH (1990) Molecular cloning of DNA from specific chromosomal regions by microdissection and sequence-independent amplification of DNA. Genomics 6:243–251

    Google Scholar 

  • Leach CR, Donald TM, Franks TK, Spiniello SS, Hanrahan CF, Timmis JN (1995) Organization and origin of a B chromosome centromeric sequence from Brachycome dichromosomatica. Chromosoma 103:708–714

    Google Scholar 

  • 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

    Google Scholar 

  • Loidl P (1988) Towards an understanding of the biological function of histone acetylation. FEBS Lett 227:91–95

    Google Scholar 

  • Maluszynska J, Heslop-Harrison JS (1991) Localization of tandemly repeated DNA sequences in Arabidopsis thaliana. Plant J 1:159–166

    Google Scholar 

  • Mole-Bajer J, Bajer AS, Zinkowski RP, Balczon RD, Brinkley BR (1990) Autoantibodies from a patient with scleroderma CREST recognized kinetochores of the higher plant Haemanthus. Proc Natl Acad Sci USA 87:3599–3603

    Google Scholar 

  • Moroi Y, Peebles C, Fritzler MJ, Steigerwald J, Tan EM (1980) Autoantibody to centromere (kinetochore) in scleroderma sera. Proc Natl Acad Sci USA 77:1627–1631

    Google Scholar 

  • Murata M, Ogura Y, Motoyoshi F (1994) Centromeric repetitive sequences in Arabidopsis thaliana. Jpn J Genet 69:361–370

    Google Scholar 

  • Murphy TD, Karpen GH (1995) Localization of centromere function in a Drosophila minichromosome. Cell 82:599–609

    Google Scholar 

  • Ouspenski II, Brinkley BR (1993) Centromeric DNA cloned from functional kinetochore fragments in mitotic cells with unreplicated genomes. J Cell Sci 105:359–367

    Google Scholar 

  • Page SL, Earnshaw WC, Choo KHA, Shaffer LG (1995) Further evidence that CENP-C is a necessary component of active centromeres: studies of a dic(X;15) with simultaneous immunofluorescence and FISH. Hum Mol Genet 4:289–294

    Google Scholar 

  • Palevitz BA (1990) Kinetochore behavior during generative cell division in Tradescantia virginiana. Protoplasma 157:120–127

    Google Scholar 

  • Pich U, Houben A, Fuchs J, Meister A, Schubert I (1994) Utility of DNA amplified by degenerate oligonucleotide-primed PCR (DOP-PCR) from the total genome and defined chromosomal regions of field beans. Mol Gen Genet 243:173–177

    Google Scholar 

  • Pluta AF, Mackay AM, Ainsztein AM, Goldberg IG, Earnshaw WC (1995) The centromere: hub of chromosomal activities. Science 270:1591–1594

    Google Scholar 

  • Schmit A-C, Stoppin V, Chevrier V, Job D, Lambert A-M (1994) Cell cycle dependent distribution of a centrosomal antigen at the perinuclear MTOC or at the kinetochores of higher plant cells. Chromosoma 103:343–351

    Google Scholar 

  • Schubert I, Rieger R (1990) Alteration by centric fission of the diploid chromosome number in Vicia faba L. Genetica 81:67–69

    Google Scholar 

  • Schubert I, Rieger R (1991) Catalogue of chromosomal and morphological mutants of Faba bean in the Gatersleben Collection, 1991. FABIS Newsl 28/29:14–22

    Google Scholar 

  • Schubert I, Dolezel J, Houben A, Scherthan H, Wanner G (1993) Refined examination of plant metaphase chromosome structure at different levels made feasible by new isolation methods. Chromosoma 102:96–101

    Google Scholar 

  • Shaw DD (1994) Centromes: moving chromosomes through space and time. Trends Ecol Evol 9:170–175

    Google Scholar 

  • Sullivan BA, Schwartz S (1995) Identification of centromeric antigens in dicentric Robertsonian translocations: CENP-C and CENP-E are necessary compoments of functional contromeres. Hum Mol Genet 4:2189–2197

    Google Scholar 

  • Sunkel CE, Coelho PA (1995) The elusive centromere: sequence divergence and functional conservation. Curr Opinion Genet Dev 5:756–767

    Google Scholar 

  • Tagle DA, Swaroop M, Lovett M, Collins FS (1993) Magnetic bead capture of expressed sequences encoded within large genomic segments. Nature 361:751–753

    Google Scholar 

  • Telenius H, Carter NP, Bebb CE, Nordenskjöld M, Ponder BAJ, Tunnacliffe A (1992) Degenerate oligonucleotide-primed PCR: general amplification of target DNA by a single degenerate primer. Genomics 13:718–725

    Google Scholar 

  • Turner BM (1993) Decoding the nucleosome. Cell 75:5–8

    Google Scholar 

  • Tyler-Smith C, Oakey RJ, Larin Z, Fisher RB, Crocker M, Affara NA, Ferguson-Smith MA, Muenke M, Zuffardi O, Jobling MA (1993) Localization of DNA sequences required for human centromere function through an analysis of rearranged Y chromosomes. Nature Genet 5:368–375

    Google Scholar 

  • van Holde KE (1988) Chromatin. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Vig BK (1994) Do specific nucleotide bases constitute the centromere? Mutat Res 309:1–10

    Google Scholar 

  • Vig BK, Latour D, Frankovich J (1994) Dissociation of minor satellite from the centromere in mouse. J Cell Sci 107:3091–3095

    Google Scholar 

  • Yen TJ, Compton DA, Wise D, Zinkowski RP, Brinkley BR, Earnshaw WC, Cleveland DW (1991) CENP-E, a novel human centromere-associated protein required for progression from metaphase to anaphase. EMBO J 10:1245–1254

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Pflanzengenetik und Kulturpflanzenforschung, Corrensstrasse 3, D-06466, Gatersleben, Germany

    A. Houben, A. Brandes, U. Pich, R. Manteuffel & I. Schubert

Authors
  1. A. Houben
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. Brandes
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. U. Pich
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. R. Manteuffel
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. I. Schubert
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by F. Mechelke

Rights and permissions

Reprints and permissions

About this article

Cite this article

Houben, A., Brandes, A., Pich, U. et al. Molecular-cytogenetic characterization of a higher plant centromere/kinetochore complex. Theoret. Appl. Genetics 93, 477–484 (1996). https://doi.org/10.1007/BF00417938

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00417938

Key words

Search

Navigation