Expression analysis of the fpr (ferredoxin-NADP+ reductase) gene in Pseudomonas putida KT2440.
The ferredoxin-NADP+ reductase (fpr) participates in cellular defense against oxidative damage. The fpr expression in Pseudomonas putida KT2440 is induced by oxidative and osmotic stresses. FinR, a LysR-type transcriptional factor near the fpr gene in the P. putida KT2440 genome, is required for induction of the fpr under both conditions. We have shown that the fpr and finR gene products can counteract the effects of oxidative and osmotic stresses. Interestingly, FinR-independent expression occurs either during a long period of incubation with paraquat or with high concentrations of oxidative stress agent. This result indicates that there may be additional regulators present in the P. putida KT2440 genome. In contrast to in vivo expression kinetics of fpr from the plant pathogen, Pseudomonas syringae, the fpr gene from P. putida KT2440 exhibited unusually prolonged expression after oxidative stress. Transcriptional fusion and Northern blot analysis studies indicated that the FinR is negatively autoregulated. Expression of the fpr promoter was higher in minimal media than in rich media during exponential phase growth. Consistent with this result, the fpr and finR mutants had a long lag phase in minimal media in contrast to wild-type growth characteristics. Antioxidants such as ascorbate could increase the growth rate of all tested strains in minimal media. This result confirmed that P. putida KT2440 experienced more oxidative stress during exponential growth in minimal media than in rich media. Endogenous promoter activity of the fpr gene is much higher during exponential growth than during stationary growth. These findings demonstrate new relationships between fpr, finR, and the physiology of oxidative stress in P. putida KT2440.
Lee
Yunho
Y
Division of Environmental Science and Ecological Engineering, Korea University, Anam-Dong 5Ga, Seungbuk-Ku, Seoul 136-701, Republic of Korea.
Peña-Llopis
Samuel
S
Kang
Yoon-Suk
YS
Shin
Hyeon-Dong
HD
Demple
Bruce
B
Madsen
Eugene L
EL
Jeon
Che Ok
CO
Park
Woojun
W
D049109:Cell Proliferation; D005287:Ferredoxin-NADP Reductase / Q000378:metabolism*; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D007700:Kinetics; D009997:Osmotic Pressure; D018384:Oxidative Stress / Q000502:physiology*; D016958:Pseudomonas putida / Q000201:enzymology* / Q000254:growth & development*
D005287:Ferredoxin-NADP Reductase