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Nucleic Acids Research, 1977, Vol. 4, No. 11 3901-3918
© 1977

Articles

Nucleosome mono-, di-, tri-, and tetramers from chicken embryo chromatin

Burghardt Wittig and Stephanie Wittig

Institut für Molekularbiologie und Biochemie, Freie Universität Berlin Arnimallee 22, D-1000 Berlin 33, GFR

Received August 24, 1977. The fractionation of gram quantities of nuclease digested chromatin from chicken embryos into nucleosome mono-, di-, tri-, and tetramers is described in detail. Each of these nucleosomal species contains a fraction soluble in 0.1 M KCl that decreases with increasing repeat number. Less histone H1 is associated with the nucleosome fractions soluble as compared to the respective fractions precipitated in 0.1 M KCl. Thermal denaturation profiles of the four nucleosomal species are monophasic. The same T of 78 °C has been determined for the KCl-soluble nucleosomes and for the KCl-insoluble monomer. The Tm of the KCl-insoluble oligomers is 79.8 °C. Multiphasic melting curves were recorded for nucleosomal material that was concentrated by lyophilisation or stored at 4 °C in 0.25 mM EDTA. Total nucleosome mono-, di-, tri-, and tetramers (consisting of both the fraction soluble and insoluble in 0.1 M KCl) have been analyzed concerning their sedimentation, diffusion, partial specific volume, and molecular weight and compared with the sedimentation and molecular weight data of KCl-soluble nucleosome mono- and tetramers.


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