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Nucleic Acids Research, 1981, Vol. 9, No. 15 3671-3680
© 1981

MOLECULAR BIOLOGY

Protection of discrete DNA fragments by the complex Hl-octamerhistones or H5-octamerhistonesafter micrococcal nuclease digestion

Serge Muyldermans, Ignace Lasters, Lode Wyns and Raymond Hamers

Laboratorium voor Algemene Biologie, Instituut voor Moleculaire Biologie V.U.B., Paardenstraat 65, B-1640 St-Genesius-Rode, Belgium

Received May 8, 1981. Several authors, including ourselves, have reported the existence of chroma-tosomes with DNA size larger than 166 bp in bird erythrocyte chromatin. Itwas tempting to correlate this increased DNA size with the presence of histoneH5.

In order to substantiate this hypothesis, we performed a micrococcal nuclease digestion kinetic on : chicken erythrocyte chromatin, either native, selectively depleted from H1, or from H1 and H5; and rat liver chromatin, either native or partially H1 depleted. The comparative analysis of the lengths of DNA in the chromatosome size region led to the following conclusions : denaturing gels clearly reveal a first discrete pause at 178 nucleotidesin H1 depleted chicken erythrocyte chromatin as well as in partially H1-depleted rat liver chromatin, before the material accumulates at thenext intermediate 166 nucleotide chromatosome pause.

the generation of all discrete chromatosome+bands is critically dependenton low ionic strength conditions and low Ca++ concentrations during the digestion, suggesting it may result from the protection of DNA cleavage sitesby histone H5 or H1, C or N terminal domains.


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