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J Bacteriol, April 1998, p. 2201-2211, Vol. 180, No. 8
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

The Bacillus subtilis DinR Binding Site: Redefinition of the Consensus Sequence

Kevin W. Winterling,1,2 David Chafin,3 Jeffery J. Hayes,3 Ji Sun,4,5 Arthur S. Levine,1 Ronald E. Yasbin,5 and Roger Woodgate1,*

Section on DNA Replication, Repair, and Mutagenesis, National Institute of Child Health and Human Development, Bethesda, Maryland 20892-27251; Department of Biological Sciences2 and Program in Molecular and Cellular Biology,4 University of Maryland, Baltimore County, Baltimore, Maryland 21228; Department of Biochemistry and Biophysics, School of Medicine and Dentistry, University of Rochester, Rochester, New York 146423; and Department of Molecular and Cell Biology, University of Texas at Dallas, Richardson, Texas 750835

Received 15 August 1997/Accepted 11 February 1998

Recently, the DinR protein was established as the cellular repressor of the SOS response in the bacterium Bacillus subtilis. It is believed that DinR functions as the repressor by binding to a consensus sequence located in the promoter region of each SOS gene. The binding site for DinR is believed to be synonymous with the formerly identified Cheo box, a region of 12 bp displaying dyad symmetry (GAAC-N4-GTTC). Electrophoretic mobility shift assays revealed that highly purified DinR does bind to such sites located upstream of the dinA, dinB, dinC, and dinR genes. Furthermore, detailed mutational analysis of the B. subtilis recA operator indicates that some nucleotides are more important than others for maintaining efficient DinR binding. For example, nucleotide substitutions immediately 5' and 3' of the Cheo box as well as those in the N4 region appear to affect DinR binding. This data, combined with computational analyses of potential binding sites in other gram-positive organisms, yields a new consensus (DinR box) of 5'-CGAACRNRYGTTYC-3'. DNA footprint analysis of the B. subtilis dinR and recA DinR boxes revealed that the DinR box is centrally located within a DNA region of 31 bp that is protected from hydroxyl radical cleavage in the presence of DinR. Furthermore, while DinR is predominantly monomeric in solution, it apparently binds to the DinR box in a dimeric state.

* Corresponding author. Mailing address: Building 6, Room 1A13, NICHD, NIH, 9000 Rockville Pike, Bethesda, MD 20892-2725. Phone: (301) 496-6175. Fax: (301) 594-1135. E-mail: woodgate{at}helix.nih.gov.




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