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Nucleic Acids Research, 1978, Vol. 5, No. 2 523-535
© 1978

Articles

Superstructure and CD spectrum as probes of chromatin integrity

Gilbert de Murcia, Gokul C. Das, Monique Erard and Michel Daune

Institut de Biologie Moléculaire et Cellulaire du CNRS, Laboratoire de Biophysique 15 rue René Descartes, 67084 Strasbourg Cédex, France

Received November 22, 1977. Two types of chromatin were extracted from the same stock of rat liver nuclei by a short exposure to micrococcal nuclease and by shearing respectively. These two materials which are identical in their protein/DNA Content and by the presence of the five histones, were compared by means of circular dichroïsm and electron microscopy. Under the electron microscope and in absence of any divalent cation a superstructure of the unfixed chromatin fiber can be viewed only with native material but is no more present in sheared one. The increase of CD signal at 280 nm (from 2000 to about 4000 cm2 deg.dmole–1 in the case of sheared chromatin is not related to the loss of superstructure but to the structural changes of DNA inside the nucleosomal core which are always produced by shearing. These two correlated observations offer new sensitive probes of the integrity of any native or reconstituted chromatin.


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