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Nitric oxide in chemostat-cultured Escherichia coli is sensed by Fnr and other global regulators: unaltered methionine biosynthesis indicates lack of S nitrosation

Nitric oxide in chemostat-cultured Escherichia coli is sensed by Fnr and other global regulators: unaltered methionine biosynthesis indicates lack of S nitrosation
Nitric oxide in chemostat-cultured Escherichia coli is sensed by Fnr and other global regulators: unaltered methionine biosynthesis indicates lack of S nitrosation

We previously elucidated the global transcriptional responses of Escherichia coli to the nitrosating agent S-nitrosoglutathione (GSNO) in both aerobic and anaerobic chemostats, demonstrated the expression of nitric oxide (NO)-protective mechanisms, and obtained evidence of critical thiol nitrosation. The present study was the first to examine the transcriptome of NO-exposed E. coli in a chemostat. Using identical conditions, we compared the GSNO stimulon with the stimulon of NO released from two NO donor compounds {3-[2-hydroxy-1-(1-methyl-ethyl)-2-nitrosohydrazino]-1-propanamine (NOC-5) and 3-(2-hydroxy-1-methyl-2-nitrosohydrazino)-N-methyl-1-propanamine (NOC-7)} simultaneously and demonstrated that there were marked differences in the transcriptional responses to these distinct nitrosative stresses. Exposure to NO did not induce met genes, suggesting that, unlike GSNO, NO does not elicit homocysteine S nitrosation and compensatory increases in methionine biosynthesis. After entry into cells, exogenous methionine provided protection from GSNO-mediated killing but not from NO-mediated killing. Anaerobic exposure to NO led to up-regulation of multiple Fnr-repressed genes and down-regulation of Fnr-activated genes, including nrfA, which encodes cytochrome c nitrite reductase, providing strong evidence that there is NO inactivation of Fnr. Other global regulators apparently affected by NO were IscR, Fur, SoxR, NsrR, and NorR. We tried to identify components of the NorR regulon by performing a microarray comparison of NO-exposed wild-type and norR mutant strains; only norVW, encoding the NO-detoxifying flavorubredoxin and its cognate reductase, were unambiguously identified. Mutation of norV or norR had no effect on E. coli survival in mouse macrophages. Thus, GSNO (a nitrosating agent) and NO have distinct cellular effects; NO more effectively interacts with global regulators that mediate adaptive responses to nitrosative stress but does not affect methionine requirements arising from homocysteine nitrosation.

Aerobiosis, Anaerobiosis, Animals, Escherichia coli, Escherichia coli Proteins, Gene Expression Regulation, Bacterial, Iron-Sulfur Proteins, Methionine, Mice, Nitric Oxide, Nitrosation, Polymerase Chain Reaction, S-Nitrosoglutathione, Trans-Activators, Journal Article, Research Support, Non-U.S. Gov't
0021-9193
1845-1855
Pullan, Steven T.
e2e1bf2a-8420-4e75-99fa-bd4283547595
Gidley, Mark D.
e05f890f-a551-44df-85d0-b860d637646d
Jones, Richard A.
2c42c836-f346-4477-b32b-764b31a34454
Barrett, Jason
34a20edd-44dd-45c9-9a76-b82fb94ee24e
Stevanin, Tania M.
f1504019-7365-4b69-8404-e9df63a5348a
Read, Robert C.
b5caca7b-0063-438a-b703-7ecbb6fc2b51
Green, Jeffrey
bd38753d-70c3-4327-9893-090ffb2b15ea
Poole, Robert K.
5f1f3b79-cf45-4ae6-89cd-e4259c03ae56
Pullan, Steven T.
e2e1bf2a-8420-4e75-99fa-bd4283547595
Gidley, Mark D.
e05f890f-a551-44df-85d0-b860d637646d
Jones, Richard A.
2c42c836-f346-4477-b32b-764b31a34454
Barrett, Jason
34a20edd-44dd-45c9-9a76-b82fb94ee24e
Stevanin, Tania M.
f1504019-7365-4b69-8404-e9df63a5348a
Read, Robert C.
b5caca7b-0063-438a-b703-7ecbb6fc2b51
Green, Jeffrey
bd38753d-70c3-4327-9893-090ffb2b15ea
Poole, Robert K.
5f1f3b79-cf45-4ae6-89cd-e4259c03ae56

Pullan, Steven T., Gidley, Mark D., Jones, Richard A., Barrett, Jason, Stevanin, Tania M., Read, Robert C., Green, Jeffrey and Poole, Robert K. (2007) Nitric oxide in chemostat-cultured Escherichia coli is sensed by Fnr and other global regulators: unaltered methionine biosynthesis indicates lack of S nitrosation. Journal of bacteriology, 189 (5), 1845-1855. (doi:10.1128/JB.01354-06).

Record type: Article

Abstract

We previously elucidated the global transcriptional responses of Escherichia coli to the nitrosating agent S-nitrosoglutathione (GSNO) in both aerobic and anaerobic chemostats, demonstrated the expression of nitric oxide (NO)-protective mechanisms, and obtained evidence of critical thiol nitrosation. The present study was the first to examine the transcriptome of NO-exposed E. coli in a chemostat. Using identical conditions, we compared the GSNO stimulon with the stimulon of NO released from two NO donor compounds {3-[2-hydroxy-1-(1-methyl-ethyl)-2-nitrosohydrazino]-1-propanamine (NOC-5) and 3-(2-hydroxy-1-methyl-2-nitrosohydrazino)-N-methyl-1-propanamine (NOC-7)} simultaneously and demonstrated that there were marked differences in the transcriptional responses to these distinct nitrosative stresses. Exposure to NO did not induce met genes, suggesting that, unlike GSNO, NO does not elicit homocysteine S nitrosation and compensatory increases in methionine biosynthesis. After entry into cells, exogenous methionine provided protection from GSNO-mediated killing but not from NO-mediated killing. Anaerobic exposure to NO led to up-regulation of multiple Fnr-repressed genes and down-regulation of Fnr-activated genes, including nrfA, which encodes cytochrome c nitrite reductase, providing strong evidence that there is NO inactivation of Fnr. Other global regulators apparently affected by NO were IscR, Fur, SoxR, NsrR, and NorR. We tried to identify components of the NorR regulon by performing a microarray comparison of NO-exposed wild-type and norR mutant strains; only norVW, encoding the NO-detoxifying flavorubredoxin and its cognate reductase, were unambiguously identified. Mutation of norV or norR had no effect on E. coli survival in mouse macrophages. Thus, GSNO (a nitrosating agent) and NO have distinct cellular effects; NO more effectively interacts with global regulators that mediate adaptive responses to nitrosative stress but does not affect methionine requirements arising from homocysteine nitrosation.

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More information

Accepted/In Press date: 5 December 2006
e-pub ahead of print date: 22 December 2006
Published date: March 2007
Keywords: Aerobiosis, Anaerobiosis, Animals, Escherichia coli, Escherichia coli Proteins, Gene Expression Regulation, Bacterial, Iron-Sulfur Proteins, Methionine, Mice, Nitric Oxide, Nitrosation, Polymerase Chain Reaction, S-Nitrosoglutathione, Trans-Activators, Journal Article, Research Support, Non-U.S. Gov't

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Local EPrints ID: 416318
URI: https://eprints.soton.ac.uk/id/eprint/416318
ISSN: 0021-9193
PURE UUID: 4f4c8729-1d44-4aa7-9f8f-0f5e949c23f1
ORCID for Robert C. Read: ORCID iD orcid.org/0000-0002-4297-6728

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Date deposited: 12 Dec 2017 17:30
Last modified: 14 Mar 2019 01:36

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Contributors

Author: Steven T. Pullan
Author: Mark D. Gidley
Author: Richard A. Jones
Author: Jason Barrett
Author: Tania M. Stevanin
Author: Robert C. Read ORCID iD
Author: Jeffrey Green
Author: Robert K. Poole

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