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J. Biochem, 1990, Vol. 107, No. 5 661-665
© 1990 Japanese Biochemical Society

research-article

Effect of Unusual Polyamines on Cleavage of DNA by Restriction Enzymes1

Hiromi Ririno, Reiko Kuwahara, Nobuko Hamasaki and Tairo Oshima2

Department of Life Science, Tokyo Institute of Technology Midori-ku, Yokohama, Kanagawa 227

2To whom all correspondence should be sent.

The effect of unusual polyamines, such as thermine, caldopentamine, caldohexamine, tris(3-aminopropyl)amine, or tetrakis(3-aminopropyl)ammonium, on the activities of various restriction endonucleases was investigated by using an Escherichia coli plasmid as a substrate, which contains a high GC content fragment from an extreme thermophile. Restriction enzymes used were Smal, BanII, Nael, Hsal, and Taql. Most of the polyamines tested were inhibitory to the enzyme activities. The larger and more branched a polyamine was, the more the activities of nucleases were inhibited. The inhibition was positively correlated with the polyamine concentration. The sites protected by a polyamine were identical to those protected by other polyamines, and also identical to those which were less sensitive to the restriction enzyme in the absence of polyamines. No sequence specificity was seen among these sites.

1 This work was partly supported by Grants-in-Aid for Scientific Research (No. 63010071, 63044047, 63124031) from the Ministry of Education, Science and Culture of Japan, and a research grant from the Japan Securities Scholarship Foundation.


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