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.
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.
Constantinidou
Chrystala
C
School of Biosciences, University of Birmingham, Birmingham B15 2TT, United Kingdom.
Hobman
Jon L
JL
Griffiths
Lesley
L
Patel
Mala D
MD
Penn
Charles W
CW
Cole
Jeffrey A
JA
Overton
Tim W
TW
D001665:Binding Sites; D047369:Chromatin Immunoprecipitation; D017931:DNA Primers / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000737:chemistry*; D004926:Escherichia coli / Q000378:metabolism / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology*; D017353:Gene Deletion; D016680:Genome, Bacterial; D005947:Glucose / Q000378:metabolism; D007506:Iron-Sulfur Proteins / Q000378:metabolism / Q000502:physiology*; D008565:Membrane Proteins / Q000737:chemistry*; D008954:Models, Biological; D009154:Mutation; D009566:Nitrates / Q000737:chemistry*; D009573:Nitrites / Q000737:chemistry*; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D010100:Oxygen / Q000378:metabolism*; D010750:Phosphoproteins / Q000737:chemistry*; D011401:Promoter Regions, Genetic; D011494:Protein Kinases / Q000737:chemistry*; D013997:Time Factors; D015533:Transcriptional Activation
D017931:DNA Primers; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C060704:FNR protein, E coli; D007506:Iron-Sulfur Proteins; D008565:Membrane Proteins; D009566:Nitrates; D009573:Nitrites; D010750:Phosphoproteins; C081466:narP protein, E coli; C088708:narQ protein, E coli; C069780:NarL protein, E coli; D011494:Protein Kinases; C074787:narX protein, E coli; D005947:Glucose; D010100:Oxygen