The flavohemoglobin of Escherichia coli confers resistance to a nitrosating agent, a "Nitric oxide Releaser," and paraquat and is essential for transcriptional responses to oxidative stress.
Escherichia coli possesses a flavohemoglobin (Hmp), product of hmp, the first microbial globin gene to be sequenced and characterized at the molecular level. Although related proteins occur in numerous prokaryotes and eukaryotic microorganisms, the function(s) of these proteins have been elusive. Here we report construction of a defined hmp mutation and its use to probe Hmp function. As anticipated from up-regulation of hmp expression by nitric oxide (NO), S-nitrosoglutathione (GSNO) or sodium nitroprusside (SNP), the hmp mutant is hypersensitive to these agents. The hmp promoter is more sensitive to SNP and S-nitroso-N-penicillamine (SNAP) than is the soxS promoter, consistent with the role of Hmp in protection from reactive nitrogen species. Additional functions for Hmp are indicated by (a) parallel sensitivity of the hmp mutant to the redox-cycling agent, paraquat, (b) inability of the mutant to up-regulate fully the soxS and sodA promoters in response to oxidative stress caused by paraquat, GSNO and SNP, and (c) failure of the mutant to accumulate reduced paraquat radical after anoxic growth. We conclude that Hmp plays a role in protection from nitrosating agents and NO-related species and oxidative stress. This protective role probably involves direct detoxification of those species and sensing of NO-related and oxidative stress.
Membrillo-Hernández
J
J
The Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, The University of Sheffield, Firth Court, Western Bank, Sheffield S10 2TN, United Kingdom.
Coopamah
M D
MD
Anjum
M F
MF
Stevanin
T M
TM
Kelly
A
A
Hughes
M N
MN
Poole
R K
RK
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004093:Dihydropteridine Reductase; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins; D006420:Hemeproteins / Q000235:genetics / Q000378:metabolism*; D016296:Mutagenesis; D009247:NADH, NADPH Oxidoreductases; D020030:Nitric Oxide Donors / Q000494:pharmacology*; D018384:Oxidative Stress; D010100:Oxygen / Q000378:metabolism; D010269:Paraquat / Q000494:pharmacology*; D013057:Spectrum Analysis; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D006420:Hemeproteins; D020030:Nitric Oxide Donors; D004093:Dihydropteridine Reductase; C076538:hmp protein, E coli; D009247:NADH, NADPH Oxidoreductases; D010269:Paraquat; D010100:Oxygen