Nuclear factor kappa B is involved in lipopolysaccharide-stimulated induction of interferon regulatory factor-1 and GAS/GAF DNA-binding in human umbilical vein endothelial cells.

Abstract

1. In this study we examined the signalling events that regulate lipopolysaccharide (LPS)-stimulated induction of interferon regulatory factor (IRF)-1 in human umbilical vein endothelial cells (HUVECs). 2. LPS stimulated a time- and concentration-dependent increase in IRF-1 protein expression, an effect that was mimicked by the cytokine, tumour necrosis factor (TNF)-alpha. 3. LPS stimulated a rapid increase in nuclear factor kappa B (NFkappaB) DNA-binding activity. Pre-incubation with the NFkappaB pathway inhibitors, N-alpha-tosyl-L-lysine chloromethyl ketone (TLCK) or pyrrolidine dithiocarbamate (PDTC), or infection with adenovirus encoding IkappaBalpha, blocked both IRF-1 induction and NFkappaB DNA-binding activity. 4. LPS and TNFalpha also stimulated a rapid activation of gamma interferon activation site/gamma interferon activation factor (GAS/GAF) DNA-binding in HUVECs. Preincubation with the Janus kinase (JAK)-2 inhibitor, AG490 blocked LPS-stimulated IRF-1 induction but did not affect GAS/GAF DNA-binding. 5. Preincubation with TLCK, PDTC or infection with IkappaBalpha adenovirus abolished LPS-stimulated GAS/GAF DNA-binding. 6. Incubation of nuclear extracts with antibodies to RelA/p50 supershifted GAS/GAF DNA-binding demonstrating the involvement of NFkappaB isoforms in the formation of the GAS/GAF complex. 7. These studies show that NFkappaB plays an important role in the regulation of IRF-1 induction in HUVECs. This is in part due to the interaction of NFkappaB isoforms with the GAS/GAF complex either directly or via an intermediate protein.

• Authors: L Liu, A Paul, C J MacKenzie, C Bryant, A Graham, R Plevin
• Journal: British journal of pharmacology (Br J Pharmacol) Vol. 134 Issue 8 Pg. 1629-38 (Dec 2001) ISSN: 0007-1188 [Print] England
• PMID: 11739238 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• DNA-Binding Proteins
• IRF1 protein, human
• Interferon Regulatory Factor-1
• Interferon-Stimulated Gene Factor 3
• Lipopolysaccharides
• NF-kappa B
• Nuclear Proteins
• Phosphoproteins
• Protein Isoforms
• Thiocarbamates
• Transcription Factors
• Tumor Necrosis Factor-alpha
• Tyrphostins
• alpha-cyano-(3,4-dihydroxy)-N-benzylcinnamide
• gamma interferon activation factor
• prolinedithiocarbamate
• Tosyllysine Chloromethyl Ketone
• Interferon-gamma
• Proline

Topics

• Adenoviridae (genetics)
• Blotting, Western
• Cells, Cultured
• DNA-Binding Proteins (biosynthesis, metabolism)
• Dose-Response Relationship, Drug
• Endothelium, Vascular (drug effects, metabolism)
• Genetic Vectors
• Humans
• Infant, Newborn
• Interferon Regulatory Factor-1
• Interferon-Stimulated Gene Factor 3
• Interferon-gamma (metabolism)
• Lipopolysaccharides (pharmacology)
• NF-kappa B (antagonists & inhibitors, metabolism)
• Nuclear Proteins (metabolism)
• Phosphoproteins (biosynthesis)
• Proline (analogs & derivatives, pharmacology)
• Protein Isoforms (metabolism)
• Thiocarbamates (pharmacology)
• Time Factors
• Tosyllysine Chloromethyl Ketone (pharmacology)
• Transcription Factors (metabolism)
• Tumor Necrosis Factor-alpha (pharmacology)
• Tyrphostins (pharmacology)
• Umbilical Veins (cytology)