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J Biol Chem, Vol. 273, Issue 7, 3895-3900, February 13, 1998
B and HIV-1 Long Terminal Repeat
Transcriptional Activation by Inducible Nitric Oxide Synthase 2 Activity
§,
From the In the human lymphoblastoid T cell line
JJhan-5.1, stably transfected with a human immunodeficiency virus-1
long terminal repeat luciferase vector, the level of luciferase
activity is dependent on activation of the nuclear factor Unité 571 du CNRS,
Bâtiment 430, Université Paris-Sud, F-91405 Orsay Cedex
and § Unité de Biologie des Rétrovirus, Institut
Pasteur, 28 rue du Dr. Roux,
F-75724 Paris Cedex 15, France
B
(NF-B) transcription factor. Tumor necrosis factor-induced
luciferase activity was not modified in JJhan-5.1 cells co-cultivated
with murine adenocarcinoma EMT-6 cells but was strongly decreased when
nitric oxide (NO) synthase 2 expression was induced in these cells. Two
NO synthase inhibitors counteracted this inhibitory effect. Tumor
necrosis factor-
binding to JJhan-5.1 cells was not modified after
incubation with EMT-6 cells. Viability and protein synthesis in
JJhan-5.1 cells were also unchanged. Induction of NF-B DNA binding
activity was inhibited when EMT-6 cells expressed NO synthase 2 activity. Aminoguanidine, which completely abolished nitrite
production, prevented this inhibition. NF-B activation was also
strongly inhibited by S-nitrosoglutathione but was
marginally affected by
N-(2-aminoethyl)-N-(2-hydroxy-2-nitrosohydrazino)-1,2-ethylenediamine. Taken together, these results indicated that NO-related species, released by EMT-6 effector cells and probably different from NO itself,
inhibited NF-B activation in human lymphoblastoid target cells.
Consequently, transcriptional activity of a long terminal repeat-driven
luciferase gene construct was markedly diminished.
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