What question did this study set out to answer?

To characterize the transcriptional regulation of the human inducible nitric oxide synthase (NOS2) gene and map its cytokine-responsive promoter regions.

February 6, 1996Proceedings of the National Academy of SciencesOpen Access

Transcriptional regulation of human inducible nitric oxide synthase (NOS2) gene by cytokines: initial analysis of the human NOS2 promoter.

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Population

Human liver epithelial cell line (AKN-1)

Comparison

Stimulation with cytokine mixture following… vs Unstimulated cells and cells transfected with…

Design

Preclinical

Follow-up

2 to 24 hours

Authors

MVMichael E. de VeraHospital de Sant Pau RSRichard A. ShapiroUniversity of Pittsburgh ANAndreas K. NüsslerUniversity of Tübingen

Discussion

Member takes

Implication

Caution extrapolating murine NOS2 data to humans; leaves open species-specific cytokine regulation for further study.

Key Points

To characterize the transcriptional regulation of the human inducible nitric oxide synthase (NOS2) gene and map its cytokine-responsive promoter regions.
Stimulated human liver epithelial AKN-1 cells with a cytokine mixture containing tumor necrosis factor alpha, interleukin 1 beta, and interferon gamma, measuring transcriptional activation by nuclear run-on assays.
Transfected cells with 5' flanking human NOS2 promoter-luciferase constructs spanning up to -16 kb, alone or fused to a heterologous thymidine kinase promoter, to measure reporter induction.
The proximal -3.8 kb region of the NOS2 promoter showed basal activity but failed to exhibit cytokine-inducible activation.
Promoter constructs extending to -5.8 kb and -7.0 kb produced a 3- to 5-fold induction of luciferase activity, while a -16 kb construct achieved a 10-fold increase after cytokine treatment.
Identified three distinct cytokine-responsive regions upstream of the human NOS2 gene (-3.8 to -5.8 kb, -5.8 to -7.0 kb, and -7.0 to -16 kb), unlike the murine promoter which requires only the proximal 1 kb.

Structured PICO

Population

Human liver epithelial cell line (AKN-1)

Intervention

Stimulation with cytokine mixture (TNF-alpha 1000 u/ml, IL-1beta 100 u/ml, IFN-gamma 250 u/ml) following transfection of various human NOS2 luciferase constructs

Comparator

Unstimulated cells and cells transfected with promoterless plasmids

Outcome

NOS2 mRNA expression, NO production (nitrite + nitrate), and NOS2 promoter activity (luciferase activity)surrogate

The human NOS2 gene is transcriptionally regulated by cytokines through multiple responsive regions located between -3.8 and -16 kb upstream, which differs significantly from the murine promoter.

Cite This Study

Vera et al. (1996) studied this question.

synapsesocial.com/papers/6a7205c131a3df824329ebfa https://doi.org/10.1073/pnas.93.3.1054

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Transcriptional regulation of human inducible nitric oxide synthase (NOS2) gene by cytokines: initial analysis of the human NOS2 promoter.

Discussion

Member takes

Implication

Key Points

Structured PICO

Cite This Study

Transcriptional regulation of human inducible nitric oxide synthase (NOS2) gene by cytokines: initial analysis of the human NOS2 promoter.

Discussion

Member takes

Implication

Key Points

Structured PICO

Cite This Study

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