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Effect of UV-Irradiation of Rat Liver Nuclei on Structural Transitions and Fractionation of the Chromatin

A. N. Prusov1 and G. Ya. Kolomijtseva1,2

1Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, 119899 Russia; fax: (095) 939-31-81.

2To whom correspondence should be addressed.

Submitted January 24, 1997; revision submitted February 28, 1997.
The effect of UV-irradiation on the chromatin fractionation from DNase I-treated rat liver nuclei by solutions with different concentration of magnesium cations and ionic strengths has been investigated. Comparative analysis of the biochemical parameters of five fractions isolated from irradiated and non-irradiated nuclei has shown marked changes in the amounts of DNA, RNA, and proteins and in the protein composition. UV-Irradiation induces the dose-dependent transition of some chromatin from the S3 and S4 fractions into the insoluble P4 fraction. Chromatin content in fractions soluble in media with high concentrations of magnesium ions (S1 and S2) does not change on irradiation. Decrease in extraction of many proteins after UV-irradiation of both nuclei and preliminarily isolated fractions of the chromatin was observed. Changes in protein content and protein electrophoresis patterns of the fractions as a consequence of exposure of the nuclei to UV-light may be the result of DNA--protein cross-linkage. A profound effect of UV-irradiation on stabilization of chromatin macrostructure in its condensed regions was observed.
KEY WORDS: UV-irradiation, fractionation, chromatin, DNA--protein cross-linking.