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Nucleic Acids Research, 1985, Vol. 13, No. 16 5747-5760
© 1985

Articles

Type II restriction endonucleases cleave single-stranded DNAs in general

Koichi Nishigaki, Yoshio Kaneko*, Hidehiko Wakuda, Yuzuru Husimi and ToyosuKe Tanaka

Department of Environmental Chemistry, Faculty of Engineering, Saitama University 255 Shimo Okubo, Urawa 338, Japan

Received June 21, 1985. Accepted July 23, 1985.

Restriction endonucleases (13 out of 18 species used for the test) were certified to cleave single-stranded(ss)DNA. Such enzymes as AvaII, Haell, Ddel, Alul, Sau3AI, AccII, TthHB8I and HapII were newly reported to cleave ssDNA. A model to account for the cleavage of ssDNA by restriction enzymes was proposed with supportive data. The essential part of the model was that restriction enzymes preferentially cleave transiently formed secondary structures (called canonical structures) in ssDNA composed of two recognition sequences with two fold rotational symmetry. This means that a restriction enzyme can cleave ssDNAs 1n general so far as the DNAs have the sequences of restriction sites for the enzyme, and that the rate of cleavage depends on the stabilities of canonical structures.

*Present address: Institute of Biotechnology, Seibu Chemical Industry Ltd., 1120 Motoaho Kamikawa-mura, Kodama-gun, Saitama Prefecture 367-02, Japan.


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