Transcription factor NNR from Paracoccus denitrificans is a sensor of both nitric oxide and oxygen: isolation of nnr* alleles encoding effector-independent proteins and evidence for a haem-based sensing mechanism

Microbiology
Yi-Ying LeeStephen Spiro

Abstract

The nitrite reductase and nitric oxide reductase regulator (NNR) from Paracoccus denitrificans activates transcription in response to nitric oxide (NO). The mechanism of NO sensing has not been elucidated for NNR, or for any of its orthologues from the FNR/CRP family of transcriptional regulators. Using regulated expression of the nnr gene in Escherichia coli, evidence has now been obtained to indicate that activation of NNR by NO does not require de novo synthesis of the NNR polypeptide. In anaerobic cultures, NNR is inactivated slowly following removal of the source of NO. In contrast, exposure of anaerobically grown cultures to oxygen causes rapid inactivation of NNR, suggesting that the protein is inactivated directly by oxygen. By random and site-directed mutagenesis, two variants of NNR were isolated (with substitutions of arginine at position 80) that show high levels of activity in anaerobic cultures in the absence of NO. These proteins remain substantially inactive in aerobic cultures, suggesting that the substitutions uncouple the NO- and oxygen-signalling mechanisms, thus providing further evidence that NNR senses both molecules. Structural modelling suggested that Arg-80 is close to the C-helix that forms the monome...Continue Reading

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Citations

Mar 20, 2012·Antioxidants & Redox Signaling·Serena RinaldoFrancesca Cutruzzolà
Mar 28, 2012·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Stephen Spiro
Jan 23, 2016·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Jason C CrackNick E Le Brun
Apr 19, 2008·Journal of Molecular Biology·Giorgio GiardinaFrancesca Cutruzzolà
Jul 3, 2010·Microbiology·Angela Hartsock, James P Shapleigh
Aug 15, 2014·Biochemical and Biophysical Research Communications·Serena RinaldoFrancesca Cutruzzolà
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Feb 23, 2007·FEMS Microbiology Reviews·Stephen Spiro
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Dec 9, 2017·Antioxidants & Redox Signaling·Jason C Crack, Nick E Le Brun
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Nov 22, 2008·Biochemical Society Transactions·Stephen Spiro
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Dec 23, 2017·FEMS Microbiology Letters·Hannah GaimsterGary Rowley
May 25, 2021·Frontiers in Molecular Biosciences·Sylvain Durand, Maude Guillier

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