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Originally published In Press as doi:10.1074/jbc.M202445200 on June 13, 2002

J. Biol. Chem., Vol. 277, Issue 36, 33148-33152, September 6, 2002
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Genetic Analysis of GalR Tetramerization in DNA Looping during Repressosome Assembly*

Mark Geanacopoulos and Sankar AdhyaDagger

From the From Laboratory of Molecular Biology, NCI, National Institutes of Health, Bethesda, Maryland 20892-4264

The Gal repressosome is a nucleoprotein complex consisting of 2 GalR dimers, 1 HU, and 1 DNA loop, which represses the transcription of the gal operon. We have adopted a structure-based genetic approach to complement ongoing physical studies of the complex. Homology-based and subsequent alanine-scanning mutageneses suggest that five residues in the DNA-distal subdomain of GalR dimer are important for repressosome formation. A further analysis of these and intragenic suppressors of looping-defective GalR mutants as well as gain-of-function mutants that permit repressosome assembly in the absence of HU show that GalR dimers contact each other in the repressosome in a partially stacked configuration.

* 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.

Dagger To whom correspondence should be addressed: Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, 37 Convent Dr., MSC 4264, Rm. 5138, Bethesda, MD 20892-4264. Tel.: 301-496-2495; Fax: 301-480-7687; E-mail: sadhya@helix.nih.gov.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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Copyright © 2002 by the American Society for Biochemistry and Molecular Biology.