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Nucleic Acids Research, 1981, Vol. 9, No. 20 5253-5268
© 1981

MOLECULAR BIOLOGY

Interaction of histone H1 with superhelical DNA. Sedimentation and electron microscopical studies at low salt concentration

Michael Böttger, Claus-Ulrich von Mickwitz, Siegfried Scherneck, Klaus Grade and Ruth Lindigkeit

Central Institute of Molecular Biology, Academy of Sciences of the GDR 1115 Berlin-Buch, GDR

Received August 3, 1981. Complexes of histone H1 with superhelical SV40 DNA obtained by direct mixing were studied in 0.1 SSC buffer corresponding to 0.02 M Na+. Depending on the molar input ratio H1/DNA three classes of sedimenting species were observed: (1) a component sedimenting similar to superhelical DNA with a sedimentation coefficient s2o, w of 25 S observable up to 335 Mol H1/Mol DNA (w/w = 2); (2) a component with s2o, w = 120 S appearing at 135 Mol H1/Mol DNA and (3) growing amounts of heterogeneous aggregates > 1000 S. Electron micrographs revealed the 25 S component to consist of double-fibers formed from one DNA molecule and the 120 S component to consist of bundles of several such double-fibers. The aggregates represent cable-like structures. The addition of ethidium bromide to 25 S complexes induces the formation of bundles, if H1 is present in a quantity which alone is not sufficient to bring about this effect. This result indicates that ethidium bromide effects a redistribution of H1 molecules and that H1 is responsible for the bundle formation.


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