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Nucleic Acids Research, 1975, Vol. 2, No. 5 667-682
© 1975

Articles

Isolation and characterization of the DNA fraction of rat liver chromatin which binds polylysine

Eugene A. Arnold, Udo Wahn and Keith E. Young

Department of Pathology, The Johns Hopkins University School of Medicine Baltimore, Maryland 21205, USA

Received February 12, 1975. The structure of eukaryotic chromatin has been investigated by isolating and analyzing the "accessible" DNA fraction of rat liver chromatin. This DNA fraction has been isolated by titrating the chromatin with the proteaseresistant D isomer of polylysine to bind the "accessible" DNA sites. After removal of chromosomal proteins by digestion with pronase, all DNA not protected from attack by bound polylysine was removed by digestion with DNase. Even after exhaustive treatment with pronase and DNase approximately 30% of the chromatin DNA remains resistant to nuclease attack. Analysis of the isolated DNA shows it to be mainly double-stranded with an average size of 200-250 base pairs. The DNA is slightly A-T rich and contains both repetitive and "single-copy" nucleotide sequences. The results suggest that there are extensive regions in chromatin where the DNA is not tightly complexed with protein. Furthermore, the DNA of these regions is similar in gross properties to the DNA of the total genome.


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