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New sites of methylcytosine-rich DNA detected on metaphase chromosomes

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Abstract

In situ immunofluorescence detection of antibodies against 5-methylcytosine on metaphase chromosomes prepared by a new procedure allows the display of new 5-methylcytosine-rich sites as compared to previously published methods. In short-term culture lymphocytes, the immunofluorescent signals give a recurrent pattern in which four types of binding sites can be distinguished. Type I sites are the secondary constrictions and a few juxtacentromeric regions, type II sites correspond to T-bands. Both types I and II sites emit a strong fluorescence. Type III sites form an R-band pattern and emit a weaker fluorescence. Type IV sites are the short arms of acrocentrics, they emit strong but polymorphic signals. The results obtained from control experiments suggest that the pattern observed is rather the expression of an uneven distribution of 5-methylcytosine-rich sites than a consequence of the various treatments used. In a lymphoblastoid cell line known to have a reduced 5-methylcytosine content, it was possible to demonstrate a heterogeneous hypomethylation among chromosome structures, principally involving type I sites. The method opens the possibility of studying in situ on chromosomes, regional variations of methylation in pathological conditions.

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Authors and Affiliations

  1. CNRS URA 147, Institut Gustave Roussy, F-94805, Villejuif Cedex, France

    Agnès Barbin

  2. CNRS URA 620, Institut Curie, 26 rue d'Ulm, 05, Paris Cedex, France

    Claire Montpellier, Anne Gibaud, Bernard Malfoy, Bernard Dutrillaux & Claire A. Bourgeois

  3. Cytogenetic Laboratory, Ljubljanska 5, 62000, Maribor, Slovenie

    Nadja Kokalj-Vokac

  4. CNRS URA 1459. Institut Pasteur, 07, Lyon Cedex, France

    Alain Niveleau

Authors
  1. Agnès Barbin
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  2. Claire Montpellier
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  3. Nadja Kokalj-Vokac
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  4. Anne Gibaud
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  5. Alain Niveleau
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  6. Bernard Malfoy
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  7. Bernard Dutrillaux
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  8. Claire A. Bourgeois
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Barbin, A., Montpellier, C., Kokalj-Vokac, N. et al. New sites of methylcytosine-rich DNA detected on metaphase chromosomes. Hum Genet 94, 684–692 (1994). https://doi.org/10.1007/BF00206964

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  • DOI: https://doi.org/10.1007/BF00206964

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