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J. Biochem, 1978, Vol. 84, No. 2 337-342
© 1978 Japanese Biochemical Society

research-article

Cellular Factors for Stimulation of Nucleosomal Template Activity for In Vitro DNA Synthesis

Hiroto AKIYOSHI and Setsuro FUJII1

Department of Enzyme Physiology, Institute for Enzyme Research School of Medicine, Tokushima University Kuramoto-cho, Tokushima, Tokushima 770

Nucleosomes isolated from Yoshida sarcoma chromatin by micrococcal nuclease treatment were relatively inactive as templates for in vitro DNA synthesis. However, the template activity increased by trypsin digestion of nucleosomes or addition of heparin to the reaction mixture. This indicates that the nucleosomal template activity is masked. A crude extract of Yoshida sarcoma cells stimulated the nucleosomal template activity. The stimulatory factor was separated into three peaks by DEAE cellulose column chromatography. The same three peaks were observed in normal rat liver extract with much lower activities, but enhanced in regenerating liver. The factors seem to stimulate DNA synthesis by activating DNA template in nucleosomes without degrading histones or changing the primary structure of nucleosomal DNA.

1Present address: Institute for Protein Research, Osaka University, Suita, Osaka 565


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