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Originally published In Press as doi:10.1074/jbc.M212462200 on January 15, 2003

J. Biol. Chem., Vol. 278, Issue 12, 10081-10086, March 21, 2003
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Regulation of the Nitric Oxide Reduction Operon (norRVW) in Escherichia coli
ROLE OF NorR AND sigma 54 IN THE NITRIC OXIDE STRESS RESPONSE*

Anne M. GardnerDagger , Christopher R. Gessner, and Paul R. Gardner

From the Division of Critical Care Medicine, Children's Hospital Medical Center, Cincinnati, Ohio 45229

Nitric oxide (NO) induces NO-detoxifying enzymes in Escherichia coli suggesting sensitive mechanisms for coordinate control of NO defense genes in response to NO stress. Exposure of E. coli to sub-micromolar NO levels under anaerobic conditions rapidly induced transcription of the NO reductase (NOR) structural genes, norV and norW, as monitored by lac gene fusions. Disruption of rpoN (sigma 54) impaired the NO-mediated induction of norV and norW transcription and NOR expression, whereas disruption of the upstream regulatory gene, norR, completely ablated NOR induction. NOR inducibility was restored to NorR null mutants by expressing NorR in trans. Furthermore, an internal deletion of the N-terminal domain of NorR activated NOR expression independent of NO exposure. Neither NorR nor sigma 54 was essential for NO-mediated induction of the NO dioxygenase (flavohemoglobin) encoded by hmp. However, elevated NOR activity inhibited NO dioxygenase induction, and, in the presence of dioxygen, NO dioxygenase inhibited norV induction by NO. The results demonstrate the role of NorR as a sigma 54-dependent regulator of norVW expression. A role for the NorR N-terminal domain as a transducer or sensor for NO is suggested.

* This work was supported by a grant from the Children's Hospital Research Foundation Trustees and Public Health Services Grant GM-65090 from the National Institutes of Health.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: ML 7006, Children's Hospital Research Foundation, 3333 Burnet Ave., Cincinnati, OH 45229. Tel.: 513-636-8712; Fax: 513-636-4892; E-mail: Anne.Gardner@cchmc.org.

Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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