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Analysis of Accessible Sites in Modified Nucleosomes

  • Chapter
Genome and Chromatin: Organization, Evolution, Function

Part of the book series: Plant Systematics and Evolution ((SYSTEMATICS,volume 2))

Abstract

Methylases from the bacterium Haemophilus influenzae are proposed as a tool for a selective modification of the internucleosomal linker DNA. The exclusive methylation at this site was demonstrated by a comparison of the rate of degradation of radioactively methylated chromatin and of total DNA with micrococcal nuclease. Analysis of the DNA fragment pattern after digestion of methylated nucleosome dimers reveals that only full size nucleosome monomers and larger fragments (e.g. dimers) are radioactively labelled, whereas smaller (subnucleosomal) DNA cleavage products do not carry any methylated portions any more. Methylation and digestion studies in the presence of polylysine support the conclusion that modification methylases and micrococcal nuclease act at the internucleosomal linker DNA, whereas polylysine interacts preferentially with the nucleosomal core DNA.—The minor groove of the nucleosomal core DNA was identified as the site of polylysine binding by competition studies with ethidium bromide, since intercalation of this molecule proceeds via the minor groove. Scatchard plots of ethidium binding parameters and analysis of nucleosome cleavage patterns indicate a competition of polylysine and ethidium bromide for binding sites in the nucleosomal core DNA.—Finally, cross-linking studies with formaldehyde and dimethylsuberi­midate suggest that the conformational change of the nucleosomal DNA due to ethidium intercalation leads to slight changes in the mode of interaction of the different nucleosomal compounds.

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Author information

Authors and Affiliations

  1. Institut für Physiologische Chemie, Philipps-Universität, Lahnberge, D-3550, Marburg/Lahn, Federal Republic of Germany

    Prof. Dr. Detlef Doenecke

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  1. Prof. Dr. Detlef Doenecke
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Editors and Affiliations

  1. Universität Kaiserslautern, Federal Republic of Germany

    Walter Nagl

  2. Institut für Biologie II, Universität Tübingen, Federal Republic of Germany

    Vera Hemleben

  3. Institut für Botanik, Universität Wien, Vienna, Austria

    Friedrich Ehrendorfer

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© 1979 Springer-Verlag

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Doenecke, D. (1979). Analysis of Accessible Sites in Modified Nucleosomes. In: Nagl, W., Hemleben, V., Ehrendorfer, F. (eds) Genome and Chromatin: Organization, Evolution, Function. Plant Systematics and Evolution, vol 2. Springer, Vienna. https://doi.org/10.1007/978-3-7091-8556-8_17

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  • DOI: https://doi.org/10.1007/978-3-7091-8556-8_17

  • Publisher Name: Springer, Vienna

  • Print ISBN: 978-3-7091-8558-2

  • Online ISBN: 978-3-7091-8556-8

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