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Journal of Bacteriology, October 2003, p. 6025-6031, Vol. 185, No. 20
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.20.6025-6031.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

DpiA Binding to the Replication Origin of Escherichia coli Plasmids and Chromosomes Destabilizes Plasmid Inheritance and Induces the Bacterial SOS Response

Christine Miller,1 Hanne Ingmer,1,2,{dagger} Line Elnif Thomsen,2 Kirsten Skarstad,3 and Stanley N. Cohen1,4*

Departments of Genetics,1 Medicine, Stanford University, Stanford, California 94305-5120,4 Department of Veterinary Microbiology, Royal Veterinary and Agricultural University, Stigboejlen 4, Frederiksberg C, DK-1870, Denmark,2 Department of Cell Biology, Institute for Cancer Research, Montebello, 0310 Oslo, Norway3

Received 23 May 2003/ Accepted 23 July 2003

The dpiA and dpiB genes of Escherichia coli, which are orthologs of genes that regulate citrate uptake and utilization in Klebsiella pneumoniae, comprise a two-component signal transduction system that can modulate the replication of and destabilize the inheritance of pSC101 and certain other plasmids. Here we show that perturbed replication and inheritance result from binding of the effector protein DpiA to A+T-rich replication origin sequences that resemble those in the K. pneumoniae promoter region targeted by the DpiA ortholog, CitB. Consistent with its ability to bind to A+T-rich origin sequences, overproduction of DpiA induced the SOS response in E. coli, suggesting that chromosomal DNA replication is affected. Bacteria that overexpressed DpiA showed an increased amount of DNA per cell and increased cell size—both also characteristic of the SOS response. Concurrent overexpression of the DNA replication initiation protein, DnaA, or the DNA helicase, DnaB—both of which act at A+T-rich replication origin sequences in the E. coli chromosome and DpiA-targeted plasmids—reversed SOS induction as well as plasmid destabilization by DpiA. Our finding that physical and functional interactions between DpiA and sites of replication initiation modulate DNA replication and plasmid inheritance suggests a mechanism by which environmental stimuli transmitted by these gene products can regulate chromosomal and plasmid dynamics.

* Corresponding author. Mailing address: Department of Genetics, Stanford University, 300 Pasteur Dr., Stanford, CA 94305-5120. Phone: (650) 723-5315. Fax: (650) 725-1536. E-mail: sncohen{at}stanford.edu.

{dagger} Present address: Department of Veterinary Microbiology, Royal Veterinary and Agricultural University, Stigboejlen 4, Frederiksberg C, DK-1870, Denmark.

Journal of Bacteriology, October 2003, p. 6025-6031, Vol. 185, No. 20
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.20.6025-6031.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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