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Originally published In Press as doi:10.1074/jbc.M005495200 on August 2, 2000

J. Biol. Chem., Vol. 275, Issue 44, 34609-34618, November 3, 2000
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The delta  Subunit of DNA Polymerase III Holoenzyme Serves as a Sliding Clamp Unloader in Escherichia coli*

Frank P. LeuDagger , Manju M. Hingorani§, Jennifer TurnerDagger ||, and Mike O'Donnell§**

From the Dagger  Joan and Sanford I. Weill Graduate School of Medical Sciences of Cornell University, Department of Pharmacology, New York, New York 10021 and § The Rockefeller University and ** Howard Hughes Medical Institute, Laboratory of DNA Replication, New York, New York 10021

In Escherichia coli, the circular beta  sliding clamp facilitates processive DNA replication by tethering the polymerase to primer-template DNA. When synthesis is complete, polymerase dissociates from beta  and DNA and cycles to a new start site, a primed template loaded with beta . DNA polymerase cycles frequently during lagging strand replication while synthesizing 1-2-kilobase Okazaki fragments. The clamps left behind remain stable on DNA (t1/2 ~ 115 min) and must be removed rapidly for reuse at numerous primed sites on the lagging strand. Here we show that delta , a single subunit of DNA polymerase III holoenzyme, opens beta  and slips it off DNA (kunloading = 0.011 s-1) at a rate similar to that of the multisubunit gamma  complex clamp loader by itself (0.015 s-1) or within polymerase (pol) III* (0.0065 s-1). Moreover, unlike gamma  complex and pol III*, delta  does not require ATP to catalyze clamp unloading. Quantitation of gamma  complex subunits (gamma , delta , delta ', chi , psi ) in E. coli cells reveals an excess of delta , free from gamma  complex and pol III*. Since pol III* and gamma  complex occur in much lower quantities and perform several DNA metabolic functions in replication and repair, the delta  subunit probably aids beta  clamp recycling during DNA replication.

* This work was supported by National Institutes of Health Grant GM 38839 (to M. O.-D.).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.

Present address: Molecular Biology and Biochemistry Dept., Wesleyan University, Middletown, CT 06459.

|| Present address: Dept. of Biochemistry and Biophysics, University of California San Francisco, San Francisco, CA 94143.

** To whom correspondence should be addressed.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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