PMID- 8798400
OWN - NLM
STAT- MEDLINE
DCOM- 19961107
LR  - 20210210
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 271
IP  - 37
DP  - 1996 Sep 13
TI  - A 3' cis-acting element is involved in tumor necrosis factor-alpha gene 
      expression in astrocytes.
PG  - 22383-90
AB  - Tumor necrosis factor-alpha (TNF-alpha) contributes to demyelinating diseases in 
      the central nervous system. Astrocytes, the major glial cells in the CNS, do not 
      constitutively express TNF-alpha, but the TNF-alpha gene is transcriptionally 
      activated in response to a variety of stimuli, including TNF-alpha itself. 
      Because of the importance of TNF-alpha in the CNS, we examined the mechanisms 
      underlying transcriptional regulation of the TNF-alpha gene in astrocytes. In 
      transient transfection assays, a plasmid construct containing 1.3 kilobase pairs 
      (kb) of 5' flanking sequence of the rat TNF-alpha gene showed high basal activity 
      that could not be further enhanced by TNF-alpha stimulation. A "marked" 10-kb 
      TNF-alpha gene construct, which contains the whole TNF-beta gene with 1.2 kb of 
      5' flanking sequence, 1.1 kb of intergenic sequence, and the whole TNF-alpha gene 
      with 3 kb of 3' flanking sequence, was able to respond to TNF-alpha stimulation. 
      Analysis of a series of 5' and 3' deletion constructs of the marked TNF-alpha 
      genes demonstrated that upstream sequence elements such as NF-kappaB are not 
      required for TNF-alpha induction and that TNF-alpha responsive elements are 
      located in the 3' flanking region of the TNF-alpha gene. We also found that a 
      TNF-alpha-inducible DNase I-hypersensitive (DH) site is present in this 3' region 
      whose deletion abolishes TNF-alpha inducibility of the marked TNF-alpha gene. 
      Electrophoresis mobility shift assays showed that TNF-alpha-inducible nuclear 
      proteins, consisting of p50 and p65 NF-kappaB proteins, specifically bind to two 
      consecutive NF-kappaB binding sites within the 3' DH site. These results indicate 
      that TNF-alpha-induced TNF-alpha gene expression in astrocytes involves p50 and 
      p65 NF-kappaB proteins binding to downstream NF-kappaB sites and concomitant 
      modulation of the chromatin structure.
FAU - Kwon, J
AU  - Kwon J
AD  - Department of Cell Biology, University of Alabama at Birmingham, Birmingham, 
      Alabama 35294-0005, USA.
FAU - Lee, S J
AU  - Lee SJ
FAU - Benveniste, E N
AU  - Benveniste EN
LA  - eng
GR  - MH-50421/MH/NIMH 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.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (NF-kappa B)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - EC 2.3.1.28 (Chloramphenicol O-Acetyltransferase)
RN  - EC 3.1.21.1 (Deoxyribonuclease I)
SB  - IM
MH  - Animals
MH  - Astrocytes/*metabolism
MH  - Base Sequence
MH  - Binding, Competitive
MH  - Cells, Cultured
MH  - Chloramphenicol O-Acetyltransferase/genetics
MH  - Deoxyribonuclease I/metabolism
MH  - Electrophoresis, Polyacrylamide Gel
MH  - *Gene Expression Regulation
MH  - Genes, Reporter
MH  - Molecular Sequence Data
MH  - NF-kappa B/metabolism
MH  - Plasmids/metabolism
MH  - Rats
MH  - *Regulatory Sequences, Nucleic Acid
MH  - Sequence Deletion
MH  - Transfection
MH  - Tumor Necrosis Factor-alpha/*genetics
EDAT- 1996/09/13 00:00
MHDA- 1996/09/13 00:01
CRDT- 1996/09/13 00:00
PHST- 1996/09/13 00:00 [pubmed]
PHST- 1996/09/13 00:01 [medline]
PHST- 1996/09/13 00:00 [entrez]
AID - S0021-9258(19)61766-7 [pii]
AID - 10.1074/jbc.271.37.22383 [doi]
PST - ppublish
SO  - J Biol Chem. 1996 Sep 13;271(37):22383-90. doi: 10.1074/jbc.271.37.22383.