Gamma-Radiation-Induced Crosslinking of Cell-Specific Chromosomal Nonhistone Protein-DNA Complexes in HeLa Chromatin

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Abstract
Specific antisera were obtained by injecting rabbits or goats with dehistonized HeLa [human cervical carcinoma] cell chromatin. Gamma irradiation (10-100 krad) of the isolated chromatin increased its immunological reactivity with these specific antisera. Conversely, irradiation of the isolated chromosomal nonhistone proteins or DNA followed by reconstitution resulted in a nearly complete loss of immunological reactivity of the reconstituted complexes. Selective protein dissociation experiments indicated that radiation crosslinked the active nonhistone proteins to the DNA. This interpretation was further substantiated by polyacrylamide gel electrophoresis of the crosslinked proteins. The immunologically specific complexes in HeLa nuclei represent a closely interacting system of chromosomal nonhistone protein(s) with DNA which may be used to study the functional structure of chromatin.

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