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Journal of Bacteriology, June 2007, p. 4410-4417, Vol. 189, No. 12
0021-9193/07/$08.00+0     doi:10.1128/JB.00080-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

The NsrR Regulon of Escherichia coli K-12 Includes Genes Encoding the Hybrid Cluster Protein and the Periplasmic, Respiratory Nitrite Reductase{triangledown}

Nina Filenko,1 Stephen Spiro,2 Douglas F. Browning,1 Derrick Squire,1 Tim W. Overton,1 Jeff Cole,1* and Chrystala Constantinidou1

School of Biosciences, University of Birmingham, Birmingham B15 2TT, United Kingdom,1 Department of Molecular and Cell Biology, University of Texas at Dallas, P.O. Box 830688, Richardson, Texas 75083-06882

Received 16 January 2007/ Accepted 10 April 2007

Successful pathogens must be able to protect themselves against reactive nitrogen species generated either as part of host defense mechanisms or as products of their own metabolism. The regulatory protein NsrR (a member of the Rrf2 family of transcription factors) plays key roles in this stress response. Microarray analysis revealed that NsrR represses nine operons encoding 20 genes in Escherichia coli MG1655, including the hmpA, ytfE, and ygbA genes that were previously shown to be regulated by NsrR. Novel NsrR targets revealed by this study include hcp-hcr (which were predicted in a recent bioinformatic study to be NsrR regulated) and the well-studied nrfA promoter that directs the expression of the periplasmic respiratory nitrite reductase. Conversely, transcription from the ydbC promoter is strongly activated by NsrR. Regulation of the nrf operon by NsrR is consistent with the ability of the periplasmic nitrite reductase to reduce nitric oxide and hence protect against reactive nitrogen species. Gel retardation assays were used to show that both FNR and NarL bind to the hcp promoter. The expression of hcp and the contiguous gene hcr is not induced by hydroxylamine. As hmpA and ytfE encode a nitric oxide reductase and a mechanism to repair iron-sulfur centers damaged by nitric oxide, the demonstration that hcp-hcr, hmpA, and ytfE are the three transcripts most tightly regulated by NsrR highlights the possibility that the hybrid cluster protein, HCP, might also be part of a defense mechanism against reactive nitrogen stress.

* Corresponding author. Mailing address: School of Biosciences, University of Birmingham, Birmingham B15 2TT, United Kingdom. Phone: 44 121 414 5440. Fax: 44 121 414 5925. E-mail: j.a.cole{at}bham.ac.uk

{triangledown} Published ahead of print on 20 April 2007.

Journal of Bacteriology, June 2007, p. 4410-4417, Vol. 189, No. 12
0021-9193/07/$08.00+0     doi:10.1128/JB.00080-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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