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Untersuchungen zur Alkylierung von Chromatin in der Rattenleber nach Gabe von 3H-Cyclophosphamid — „Feinverteilung“ und Kinetik

Investigation of alkylation in rat liver chromatin after application of 3H-cyclophosphamid — “fine-distribution” and kinetics

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Summary

The alkylation of chromatin constituents (DNA, histones and nonhistones) in liver cell nuclei was investigated at various times after intraperitoneal injection of rats with 3H-cyclophosphamid. The highest alkylation was found in the DNA, the lowest in the histones; the euchromatic portions were alkylated several times higher compared to those of the heterochromatin. The eventual elimination of 3H-activity with time indicates that cyclophosphamid leads to repair processes in the DNA, this conclusion being supported by other experimental observations. In some of the 16 subfractions of the nonhistone proteins, alkylated portions are apparantly eliminated by normal protein turnover; however, in other nonhistones this elimination is inhibited, whereby in one subfraction it seems to be accelerated. The results of analog experiments with 14C-tryptophan as a precursor for nonhistone protein synthesis serve as a reference for this. In supplementary in-vitro model experiments using triaziquon as an alkylating agent indications for DNA-Protein-cross-links in chromatin could be obtained by the technique of X-ray low angle scattering, and according to sedimentation behaviour.

Zusammenfassung

Zu verschiedenen Zeiten nach der i.p. Verabfolgung von 3H-Cyclophosphamid an Ratten wurde (über die Bestimmung des 3H-Gehaltes) die Alkylierung der Chromatinbausteine DNA, Histone und Nichthistonproteine in den Zellkernen der Leber untersucht. Die höchste Alkylierung fand sich in der DNA, die geringste in den Histonen; die euchromatischen Anteile wurden um ein Vielfaches stärker alkyliert als die heterochromatischen. Der zeitliche Verlauf der Eliminierung der 3H-Aktivität weist, ergänzt durch andere Befunde, darauf hin, daß es nach Einwirkung von Cyclophosphamid an der DNA zu Repariervorgängen kommt. Bei 16 untersuchten Unterfraktionen der Nichthistone werden die alkylierten Anteile zum Teil offenbar über den normalen Proteinumsatz eliminiert; diese Eliminierung ist bei einigen der Nichthistone jedoch gehemmt, dagegen bei einer Unterfraktion anscheinend beschleunigt. Als Bezugssystem für diese Untersuchungen dienten die Ergebnisse analoger Versuche mit 14C-Tryptophan als Vorstufe der Nichthistonproteine. Bei ergänzenden in-vitro-Modellversuchen mit Triaziquon als Alkylans konnten mit Hilfe der Röntgenkleinwinkelstreuungstechnik und aufgrund des Sedimentationsverhaltens Hinweise für das Vorliegen von DNA-Protein-Vernetzungen im Chromatin gewonnen werden.

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Author notes

  1. K. Kruse

    Present address: Abteilung für allgemeine Pädiatrie im Fachbereich Medizin der Christian-Albrechts-Universität, Fröbelstr. 15/17, D-2300, Kiel

Authors and Affiliations

  1. Abteilung für Medizinische Molekularbiologie im Klinisch-Theoretischen Zentrum der Medizinischen Hochschule Lübeck, Ratzeburger Allee 160, D-2400, Lübeck 1

    E. Harbers, P. Warnecke, H. Hollandt & K. Kruse

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  1. E. Harbers
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  2. P. Warnecke
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  3. H. Hollandt
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  4. K. Kruse
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Harbers, E., Warnecke, P., Hollandt, H. et al. Untersuchungen zur Alkylierung von Chromatin in der Rattenleber nach Gabe von 3H-Cyclophosphamid — „Feinverteilung“ und Kinetik. Z. Krebsforsch. 88, 237–254 (1977). https://doi.org/10.1007/BF00305362

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