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Originally published In Press as doi:10.1074/jbc.M101694200 on April 20, 2001

J. Biol. Chem., Vol. 276, Issue 26, 23581-23588, June 29, 2001
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Sau3AI, a Monomeric Type II Restriction Endonuclease That Dimerizes on the DNA and Thereby Induces DNA Loops*

Peter FriedhoffDagger , Rudi Lurz§, Gerhild Lüder§, and Alfred PingoudDagger

From the Dagger  Institut für Biochemie, Justus-Liebig-Universität Giessen, D-35392 Giessen, Germany and the § Max-Planck-Institut für Molekulare Genetik, Ihnestrasse 73, D-14195 Berlin-Dahlem, Germany

Here, we report that Sau3AI, an unusually large type II restriction enzyme with sequence homology to the mismatch repair protein MutH, is a monomeric enzyme as shown by gel filtration and ultracentrifugation. Structural similarities in the N- and C-terminal halves of the protein suggest that Sau3AI is a pseudo-dimer, i.e. a polypeptide with two similar domains. Since Sau3AI displays a nonlinear dependence of cleavage activity on enzyme concentration and a strong preference for substrates with two recognition sites over those with only one, it is likely that the functionally active form of Sau3AI is a dimer of a pseudo-dimer. Indeed, electron microscopy studies demonstrate that two distant recognition sites are brought together through DNA looping induced by the simultaneous binding of two Sau3AI molecules to the DNA. We suggest that the dimeric form of Sau3AI supplies two DNA-binding sites, one that is associated with the catalytic center and one that serves as an effector site.

* This work was supported by Deutsche Forschungsgemeinschaft Grant Pi-122/12-3 and the Dr. Herbert Stolzenberg Stiftung.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Inst. für Biochemie (FB8), Justus-Liebig-Universität, Heinrich-Buff-Ring 58, D-35392 Giessen, Germany. Tel.: 49-641-99-35400; Fax: 49-641-99-35409; E-mail: Alfred.M.Pingoud@chemie.bio.uni-giessen.de.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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Copyright © 2001 by the American Society for Biochemistry and Molecular Biology.