PMID- 14581470
OWN - NLM
STAT- MEDLINE
DCOM- 20040210
LR  - 20211203
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 279
IP  - 2
DP  - 2004 Jan 9
TI  - Tumor necrosis factor-alpha inhibits endothelial nitric-oxide synthase gene 
      promoter activity in bovine aortic endothelial cells.
PG  - 963-9
AB  - 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.
FAU - Anderson, Hope D I
AU  - Anderson HD
AD  - Diabetes Center and Department of Medicine, University of California at San 
      Francisco, San Francisco, California 94143-0540, USA.
FAU - Rahmutula, Dolkun
AU  - Rahmutula D
FAU - Gardner, David G
AU  - Gardner DG
LA  - eng
GR  - HL45637/HL/NHLBI NIH HHS/United States
GR  - P30 DK63720/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
DEP - 20031027
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (I-kappa B Proteins)
RN  - 0 (NFKBIA protein, human)
RN  - 0 (RNA, Messenger)
RN  - 0 (SP3 protein, human)
RN  - 0 (Sp1 Transcription Factor)
RN  - 0 (Transcription Factors)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 139874-52-5 (NF-KappaB Inhibitor alpha)
RN  - 148710-94-5 (Sp3 Transcription Factor)
RN  - EC 1.13.12.- (Luciferases)
RN  - EC 1.14.13.39 (NOS3 protein, human)
RN  - EC 1.14.13.39 (Nitric Oxide Synthase)
RN  - EC 1.14.13.39 (Nitric Oxide Synthase Type III)
RN  - EC 2.7.11.1 (Protein Serine-Threonine Kinases)
RN  - EC 2.7.11.10 (CHUK protein, human)
RN  - EC 2.7.11.10 (I-kappa B Kinase)
RN  - EC 2.7.11.10 (IKBKB protein, human)
RN  - EC 2.7.11.10 (IKBKE protein, human)
SB  - IM
MH  - Animals
MH  - Aorta/*cytology
MH  - Blotting, Western
MH  - Cattle
MH  - Cell Nucleus/metabolism
MH  - DNA-Binding Proteins/metabolism
MH  - Dose-Response Relationship, Drug
MH  - Endothelial Cells/cytology
MH  - Endothelium, Vascular/*cytology
MH  - Genes, Dominant
MH  - Genes, Reporter
MH  - Humans
MH  - I-kappa B Kinase
MH  - I-kappa B Proteins/metabolism
MH  - Luciferases/metabolism
MH  - Mutation
MH  - NF-KappaB Inhibitor alpha
MH  - Nitric Oxide Synthase/*antagonists & inhibitors/*genetics
MH  - Nitric Oxide Synthase Type III
MH  - Phosphorylation
MH  - Plasmids/metabolism
MH  - *Promoter Regions, Genetic
MH  - Protein Binding
MH  - Protein Serine-Threonine Kinases/metabolism
MH  - RNA Processing, Post-Transcriptional
MH  - RNA, Messenger/metabolism
MH  - Sp1 Transcription Factor/metabolism
MH  - Sp3 Transcription Factor
MH  - Time Factors
MH  - Transcription Factors/metabolism
MH  - Transfection
MH  - Tumor Necrosis Factor-alpha/metabolism/*physiology
EDAT- 2003/10/29 05:00
MHDA- 2004/02/11 05:00
CRDT- 2003/10/29 05:00
PHST- 2003/10/29 05:00 [pubmed]
PHST- 2004/02/11 05:00 [medline]
PHST- 2003/10/29 05:00 [entrez]
AID - S0021-9258(18)52705-8 [pii]
AID - 10.1074/jbc.M309552200 [doi]
PST - ppublish
SO  - J Biol Chem. 2004 Jan 9;279(2):963-9. doi: 10.1074/jbc.M309552200. Epub 2003 Oct 
      27.