Role of protein kinase A in LPS-induced activation of NF-kappa B proteins of a mouse macrophage-like cell line, J774.
The electrophoretic mobility shift assays (EMSA) with the use of the synthetic HIV-1 NF-kappa B motif as a probe, showed that LPS-treatment of J774 cells (a mouse macrophage cell line) leads to the activation of the fast-moving (denoted as B1) and the slow-moving NF-kappa B (denoted as B2). The binding of both B1 and B2 to the NF-kappa B probe was inhibited specifically by either unlabelled NF-kappa B, or competitor probes, but not by unrelated probes. LPS-induced activation of NF-kappa B was inhibited by a protein kinase A (PKA) inhibitor (H-89), but not by a protein kinase C (PKC) inhibitor (H-7). PMA itself failed to activate NF-kappa B and the depletion of PKC did not prevent LPS-induced activation of NF-kappa B. The pre-treatment of J774 cells with dibutyric cAMP, forskolin, prostaglandin E2 or cholera toxin resulted in NF-kappa B activation. Thus, these data suggested a probable involvement of PKA in LPS-induced NF-kappa B activation in macrophages.
Muroi
M
M
Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City 66160-7420.
Suzuki
T
T
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003994:Bucladesine / Q000494:pharmacology; D002772:Cholera Toxin / Q000494:pharmacology; D005576:Colforsin / Q000494:pharmacology; D016384:Consensus Sequence; D000242:Cyclic AMP / Q000502:physiology*; D015232:Dinoprostone / Q000494:pharmacology; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D007546:Isoquinolines / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D008262:Macrophage Activation / Q000187:drug effects*; D008264:Macrophages / Q000187:drug effects* / Q000201:enzymology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000378:metabolism; D010766:Phosphorylation; D010879:Piperazines / Q000494:pharmacology; D011494:Protein Kinases / Q000502:physiology*; D011499:Protein Processing, Post-Translational; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013449:Sulfonamides; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D051841:Transcription Factor RelB; D014157:Transcription Factors; D014158:Transcription, Genetic / Q000187:drug effects; D014407:Tumor Cells, Cultured
D007546:Isoquinolines; D008070:Lipopolysaccharides; D016328:NF-kappa B; D009363:Neoplasm Proteins; D010879:Piperazines; D011518:Proto-Oncogene Proteins; C497594:Relb protein, mouse; D013449:Sulfonamides; D014157:Transcription Factors; D051841:Transcription Factor RelB; D005576:Colforsin; D003994:Bucladesine; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D002772:Cholera Toxin; D000242:Cyclic AMP; D011494:Protein Kinases; D015232:Dinoprostone; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide; D013755:Tetradecanoylphorbol Acetate