Regulation of the human tumor necrosis factor-alpha promoter by angiotensin II and lipopolysaccharide in cardiac fibroblasts: different cis-acting promoter sequences and transcriptional factors.

Abstract

We recently showed that angiotensin (ANG) II as well as mechanical stretch stimulated production of tumor necrosis factor (TNF) in cardiac fibroblasts. Presently, we examined the molecular mechanisms by which ANGII and lipopolysaccharide (LPS) upregulate TNF-alpha gene expression. In neonatal rat cardiac fibroblasts, increased transcription of TNF-alpha mRNA was detected as luciferase activity associated with activity of the TNF-alpha promoter. Progressive deletion from this promoter located the LPS-responsive region between -200 and -120 bp from the transcription initiation site, while the sequence between -120 and -70 bp was required for ANGII-induced expression. Next, we examined which cis-acting sequences in the TNF-alpha promoter region were essential for induction of TNF-alpha transcription. Competition analysis by electrophoretic mobility shift assay with and without specific antibodies showed that LPS increased binding of Sp1 and Sp3 to the Sp1-binding site, while Egr-1 was unimportant. With ANGII, binding of ATF-2/c-jun to the CRE site was required for TNF-alpha gene induction; neither Ets nor NF-kappaB was essential. Mutation analysis confirmed that response to LPS relied upon the Sp1 site in the TNF-alpha promoter, while the CRE-binding site was essential for stimulation by ANGII. We concluded that since TNF-alpha gene expression is transcriptionally activated by ANGII or LPS in cardiac fibroblasts via different cis-acting sequences in the TNF-alpha promoter and different transcriptional factors, mechanisms inducing TNF production differ between heart failure or cardiac hypertrophy and infectious disease.

• Authors: Hiroko Sato, Atai Watanabe, Toru Tanaka, Norimichi Koitabashi, Masashi Arai, Masahiko Kurabayashi, Tomoyuki Yokoyama
• Journal: Journal of molecular and cellular cardiology (J Mol Cell Cardiol) Vol. 35 Issue 10 Pg. 1197-205 (Oct 2003) ISSN: 0022-2828 [Print] England
• PMID: 14519430 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• DNA-Binding Proteins
• Lipopolysaccharides
• RNA, Messenger
• SP3 protein, human
• Sp1 Transcription Factor
• Sp3 protein, rat
• Transcription Factors
• Tumor Necrosis Factor-alpha
• Sp3 Transcription Factor
• Luciferases

Topics

• Animals
• Animals, Newborn
• Base Sequence
• Binding Sites
• Cell Nucleus (metabolism)
• Cells, Cultured
• DNA Mutational Analysis
• DNA-Binding Proteins (metabolism)
• Fibroblasts (metabolism)
• Gene Deletion
• Gene Expression Regulation
• Humans
• Lipopolysaccharides (metabolism)
• Luciferases (metabolism)
• Molecular Sequence Data
• Mutation
• Myocardium (metabolism)
• Plasmids (metabolism)
• Promoter Regions, Genetic
• Protein Binding
• RNA, Messenger (metabolism)
• Rats
• Sp1 Transcription Factor (metabolism)
• Sp3 Transcription Factor
• Transcription Factors (metabolism)
• Transcription, Genetic
• Transcriptional Activation
• Tumor Necrosis Factor-alpha (genetics)
• Up-Regulation