Involvement of diacylglycerol production in activation of nuclear factor kappaB by a CD14-mediated lipopolysaccharide stimulus.
Exposure of Chinese hamster CHO-K1 transfectant cells expressing mouse CD14 (CHO/CD14 cells) to lipopolysaccharide (LPS) induced rapid elevation of the cellular diacylglycerol (DAG) and choline/phosphocholine levels and nuclear translocation of nuclear factor kappaB (NFkappaB). When cells were incubated with short-chain DAG analogues or bacterial phospholipase C, NFkappaB activation occurred even without the LPS stimulus. Treatment of CHO/CD14 cells with tricyclo[5.2.1.0(2.6)]decyl-(9[8])xanthogenate (D609), an inhibitor of phosphatidylcholine-specific phospholipase C and phospholipase D, almost completely inhibited not only the LPS-dependent production of DAG and choline/phosphocholine but also the LPS-dependent NFkappaB activation. In contrast, treatment of cells with 1-(6-{[3-methoxyoestra-1,3, 5(10)-trien-17beta-yl]-1H-pyrrole-2,5-dione (U73122), an inhibitor of phosphatidylinositol-specific phospholipase C in vitro, did not affect the LPS-dependent activation of NFkappaB. Production of DAG and activation of NFkappaB after the LPS stimulus were observed in mouse macrophage-like J774.1 cells, and this response to LPS by J774. 1 cells was also inhibited by D609. These results suggest that the production of DAG from phosphatidylcholine was upstream of NFkappaB activation in response to a CD14-mediated LPS stimulus.
Yamamoto
H
H
Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162, Japan.
Hanada
K
K
Nishijima
M
M
D000818:Animals; D001665:Binding Sites; D001952:Bridged-Ring Compounds / Q000494:pharmacology*; D016466:CHO Cells; D006224:Cricetinae; D004075:Diglycerides / Q000378:metabolism*; D004926:Escherichia coli; D007700:Kinetics; D018950:Lipopolysaccharide Receptors / Q000096:biosynthesis / Q000502:physiology*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008264:Macrophages; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism*; D009636:Norbornanes; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology*; D010739:Phospholipase D / Q000037:antagonists & inhibitors; D010767:Phosphorylcholine / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis; D013859:Thiocarbamates; D013871:Thiones / Q000494:pharmacology*; D014162:Transfection; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors
D001952:Bridged-Ring Compounds; D004075:Diglycerides; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D016328:NF-kappa B; D009636:Norbornanes; D009838:Oligodeoxyribonucleotides; D010726:Phosphodiesterase Inhibitors; D011994:Recombinant Proteins; D013859:Thiocarbamates; D013871:Thiones; D010767:Phosphorylcholine; C046498:tricyclodecane-9-yl-xanthogenate; D010738:Type C Phospholipases; D010739:Phospholipase D