Cited by 255Cytokine Signaling Pathways and InteractionsNF-κB Signaling PathwaysNitric Oxide and Endothelin Effects

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Signal transducers and activators of transcription 3 (STAT3) inhibits transcription of the inducible nitric oxide synthase gene by interacting with nuclear factor κB

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YU, Zhiyuan, ZHANG, Wenzheng, KONE, Bruce C.·2002·Biochemical Journal

DOI 10.1042/bj20020588PMC PMC1222853

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Abstract

Prolific generation of NO by inducible nitric oxide synthase (iNOS) can cause unintended injury to host cells during glomerulonephritis and other inflammatory diseases. While much is known about the mechanisms of iNOS induction, few transcriptional repressors have been found. We explored the role of signal transducers and activators of transcription 3 (STAT3) proteins in interleukin (IL)-1β- and lipopolysaccharide (LPS)+interferon (IFN)-γ-mediated iNOS induction in murine mesangial cells. Both stimuli induced rapid phosphorylation of STAT3 and sequence-specific STAT3 DNA-binding activity. Supershift assays with a STAT3 element probe demonstrated that nuclear factor κB (NF-κB) p65 and p50 complexed with STAT3 in the DNA—protein complex. The direct interaction of STAT3 and NF-κB p65 was verified in vivo by co-immunoprecipitation and in vitro by pull-down assays with glutathione S-transferase-NF-κB p65 fusion protein and in vitro-translated STAT3α. Overexpression of STAT3 dramatically inhibited IL-1β- or LPS+IFN-γ-mediated induction of iNOS promoter-luciferase constructs that contained the wild-type iNOS promoter or ones harbouring mutated STAT-binding elements. In tests of indirect inhibitory effects of STAT3, overexpression of STAT3 dramatically inhibited the activity of an NF-κB-dependent promoter devoid of STAT-binding elements without affecting NF-κB DNA-binding activity. Thus STAT3, via direct interactions with NF-κB p65, serves as a dominant-negative inhibitor of NF-κB activity to suppress indirectly cytokine induction of the iNOS promoter in mesangial cells. These results provide a new model for the termination of NO production by activated iNOS following exposure to pro-inflammatory stimuli.

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Authors: YU, Z., ZHANG, W., & KONE, B. C.
Affiliations: The University of Texas Health Science Center at Houston (US)
ORCID: YU, Zhiyuan
ZHANG, Wenzheng
Venue: Biochemical Journal
Publisher: Portland Press Ltd.
Published: 2002-10-01
Volume: 367
Issue: 1
Pages: 97-105
ISSN: 0264-6021 (ISSN-L)
1470-8728
DOI: 10.1042/bj20020588
PMC: PMC1222853
OpenAlex: W1983667136
Cited by: 255
References: 35
Citations by year: 2026: 2 · 2025: 2 · 2024: 4 · 2023: 7 · 2022: 8 · 2021: 10 · 2020: 7 · 2019: 6 · 2018: 8 · 2017: 20 · 2016: 13 · 2015: 13 · 2014: 19 · 2013: 16 · 2012: 14
FWCI: 4.29
Type: journal article
Language: en
Open access: bronze
doi.org
Locations: Biochemical Journal (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
Funders: National Institutes of Health (5r01dk050745-05)
National Institute of General Medical Sciences (P50GM038529, P50 GM 38529)
National Institute of Diabetes and Digestive and Kidney Diseases (R01 DK 50745)
Topics: Life Sciences › Immunology and Microbiology › Immunology › Cytokine Signaling Pathways and Interactions
Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › NF-κB Signaling Pathways
Health Sciences › Medicine › Physiology (medical) › Nitric Oxide and Endothelin Effects
Keywords: STAT3, inducible nitric oxide synthase, NF-κB p65, mesangial cells, IL-1β, lipopolysaccharide, interferon-gamma, nitric oxide production, glomerulonephritis, transcriptional repressor, dominant negative inhibition
MeSH terms: Animals, Cell Nucleus, Cells, Cultured, DNA, DNA-Binding Proteins, Electrophoresis, Polyacrylamide Gel, Genes, Dominant, Glutathione Transferase, Inflammation, Interleukin-1, Nitric Oxide, Nitrites, Phosphorylation, Plasmids, Precipitin Tests, Promoter Regions, Genetic, Protein Binding, Recombinant Fusion Proteins, Transcription, Genetic, Transfection, Protein Biosynthesis, Blotting, Western, Trans-Activators, Cytokines, Mutagenesis, Site-Directed, NF-kappa B, Nitric Oxide Synthase, Protein Isoforms, STAT3 Transcription Factor, Mice, Transcription Factor RelA, NF-kappa B p50 Subunit, Nitric Oxide Synthase Type II

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Signal transducers and activators of transcription 3 (STAT3) inhibits transcription of the inducible nitric oxide synthase gene by interacting with nuclear factor κB

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