Tumor Necrosis Factor-α Inhibits Endothelial Nitric-oxide Synthase Gene Promoter Activity in Bovine Aortic Endothelial Cells
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Tumor necrosis factor-alpha (TNF-alpha) has been shown to reduce endothelial nitric-oxide synthase (eNOS) gene expression through post-transcriptional regulation of mRNA stability. The current study documented an independent effect of the cytokine on the eNOS gene promoter. TNF-alpha effected a time- and dose-dependent reduction in activity of a transiently transfected human -1197 eNOS-luciferase reporter. This reduction was inhibited by co-transfection of dominant negative IKKbeta as well as a nonphosphorylatable constitutively suppressive mutant of IkappaB implying involvement of the NFkappaB cascade in the inhibitory effect. The locus of the TNF-alpha-dependent inhibition was traced to two Sp1-binding sites positioned between -109 and -95 and -81 and -67 relative to the transcription start site. Electrophoretic mobility shift analysis and immunoperturbation studies showed evidence for Sp1 and Sp3 binding to each element. TNF-alpha treatment had no effect on the binding pattern to the downstream (-81 to -67) site but did suppress association of Sp1 and Sp3 to the upstream (-109 to -95) site. Collectively, these data indicate that TNF-alpha exerts transcriptional, as well as post-transcriptional, effects on eNOS gene expression and suggest a potential mechanism to account for the endothelial dysfunction that accompanies disorders such as diabetes mellitus and heart failure.
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TNFα downregulates NOS3 expression and abundance by decreasing the stability of NOS3 mRNA, effectively shortening its half-life.
“was shown to downregulate NOS3 expression and abundance [123, 131-134] by decreasing the stability of NOS3 mRNA [135, 136], effectively shortening its half-life [137].”
From Regulation of obesity and insulin resistance by nitric oxide (2014) · cites this paper as [134]
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- Authors
- Anderson, H. D. I., Rahmutula, D., & Gardner, D. G.
- Affiliations
- University of California, San Francisco (US)
- Corresponding author
- Gardner, David G.
- ORCID
- Anderson, Hope D.I.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2004-01
- Volume
- 279
- Issue
- 2
- Pages
- 963-969
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W1984870795
- Cited by
- 186
- References
- 69
- Citations by year
- 2026: 6 · 2025: 4 · 2024: 6 · 2023: 4 · 2022: 11 · 2021: 4 · 2020: 6 · 2019: 3 · 2018: 3 · 2017: 7 · 2016: 11 · 2015: 2 · 2014: 10 · 2013: 8 · 2012: 10
- FWCI
- 8.43
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Funders
- National Heart, Lung, and Blood Institute (HL-45637, R01HL045637)
- National Institute of Diabetes and Digestive and Kidney Diseases (P30-DK-63720, P30DK063720)
- Topics
- Health Sciences › Medicine › Physiology › Nitric Oxide and Endothelin Effects
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › NF-κB Signaling Pathways
- Health Sciences › Medicine › Immunology and Allergy › Cell Adhesion Molecules Research
- Keywords
- TNF-α, endothelial nitric oxide synthase, transcriptional regulation, Sp1 binding site, Sp3, NF-κB signaling, IκB, IKKβ, endothelial dysfunction, bovine aortic endothelial cells, luciferase reporter
- MeSH terms
- NF-KappaB Inhibitor alpha, Animals, Aorta, Cattle, Cell Nucleus, DNA-Binding Proteins, Dose-Response Relationship, Drug, Endothelium, Vascular, Genes, Dominant, Humans, Luciferases, Mutation, Phosphorylation, Plasmids, Promoter Regions, Genetic, Protein Binding, RNA Processing, Post-Transcriptional, RNA, Messenger, Time Factors, Transcription Factors, Transfection, Tumor Necrosis Factor-alpha, Blotting, Western, Sp1 Transcription Factor, Protein Serine-Threonine Kinases, Genes, Reporter, Nitric Oxide Synthase, I-kappa B Proteins, Endothelial Cells, I-kappa B Kinase, Sp3 Transcription Factor, Nitric Oxide Synthase Type III
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