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Originally published In Press as doi:10.1074/jbc.M512312200 on December 22, 2005

J. Biol. Chem., Vol. 281, Issue 8, 4802-4815, February 24, 2006
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A Reassessment of the FNR Regulon and Transcriptomic Analysis of the Effects of Nitrate, Nitrite, NarXL, and NarQP as Escherichia coli K12 Adapts from Aerobic to Anaerobic Growth*Formula

Chrystala Constantinidou, Jon L. Hobman, Lesley Griffiths, Mala D. Patel, Charles W. Penn, Jeffrey A. Cole, and Tim W. Overton1

From the School of Biosciences, University of Birmingham, Birmingham B15 2TT, United Kingdom

The transcription factor FNR, the regulator of fumarate and nitrate reduction, regulates major changes as Escherichia coli adapts from aerobic to anaerobic growth. In an anaerobic glycerol/trimethylamine N-oxide/fumarate medium, the fnr mutant grew as well as the parental strain, E. coli K12 MG1655, enabling us to reveal the response to oxygen, nitrate, and nitrite in the absence of glucose repression or artifacts because of variations in growth rate. Hence, many of the discrepancies between previous microarray studies of the E. coli FNR regulon were resolved. The current microarray data confirmed 31 of the previously characterized FNR-regulated operons. Forty four operons not previously known to be included in the FNR regulon were activated by FNR, and a further 28 operons appeared to be repressed. For each of these operons, a match to the consensus FNR-binding site sequence was identified. The FNR regulon therefore minimally includes at least 103, and possibly as many as 115, operons. Comparison of transcripts in the parental strain and a narXL deletion mutant revealed that transcription of 51 operons is activated, directly or indirectly, by NarL, and a further 41 operons are repressed. The narP gene was also deleted from the narXL mutant to reveal the extent of regulation by phosphorylated NarP. Fourteen promoters were more active in the narP+ strain than in the mutant, and a further 37 were strongly repressed. This is the first report that NarP might function as a global repressor as well as a transcription activator. The data also revealed possible new defense mechanisms against reactive nitrogen species.


Received for publication, November 16, 2005 , and in revised form, December 20, 2005.

* This work was supported by the UK Biotechnology and Biological Sciences Research Council Grants JIF13209, P20180 [GenBank] (to J. A. C.), and EGA16107. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Formula The on-line version of this article (available at http://www.jbc.org) contains Tables S1 and S2.

1 To whom correspondence should be addressed: School of Biosciences, University of Birmingham, Birmingham B15 2TT, UK. Tel.: 44-121-414-5435; Fax: 44-121-414-5925; E-mail: t.w.overton{at}bham.ac.uk.


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