title: High affinity receptor for IgE stimulation activates protein kinase D augmenting activator protein-1 activity for cytokine producing in mast cells.
abstract: Protein kinase D (PKD) is a serine-threonine kinase involved in the activation of a variety of cells. In mast cells, activation of PKD by cross-linking of high affinity receptor for IgE (FcepsilonRI) has been reported, but little is known for its effects on cytokine production. We investigated the roles of PKD on FcepsilonRI-induced activator protein-1 (AP-1) activation and proinflammatory cytokine productions in mast cells. Pharmacological inhibition of PKD strongly inhibited production of interleukin (IL)-13 and tumor necrosis factor (TNF)-alpha induced by FcepsilonRI stimulation, and the overexpression of PKD significantly increased the IL-13 and TNF-alpha production. Reporter assay revealed that the overexpression of PKD enhanced FcepsilonRI-induced IL-13 promoter activation, and that the 5'-flanking region of IL-13 gene from positions -110 to -52 was under the regulation of PKD. The overexpression of PKD enhanced the induction of AP-1 luciferase activity by FcepsilonRI stimulation, while it had no effect on luciferase activities dependent upon NF-kappaB and NF-AT activated by FcepsilonRI stimulation. In EMSA, c-Jun and c-Fos appear to be the major components of AP-1 complexes activated by FcepsilonRI stimulation. Moreover the overexpression of PKD strongly enhanced the phosphorylation of both c-Jun and c-Fos following FcepsilonRI stimulation. Although stress-activated protein kinase/c-Jun N-terminal kinase (JNK) is known to be an important regulator for c-Jun phosphorylation and AP-1 activation, overexpression and inhibition of PKD had no effects on JNK phosphorylation. These results suggest that PKD may play a pivotal role in FcepsilonRI-induced cytokine production in mast cells through the activation of c-Jun, c-Fos, and AP-1.
authors.affiliation: Department of Molecular Cell Immunology and Allergology, Nihon University School of Medical Science, Tokyo, Japan.
authors.forename: Kyoko
authors.initials: K
authors.lastname: Yamashita
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Gon
authors.forename: Toshibumi
authors.initials: T
authors.lastname: Shimokawa
authors.forename: Satoshi
authors.initials: S
authors.lastname: Nunomura
authors.forename: Daisuke
authors.initials: D
authors.lastname: Endo
authors.forename: Naoko
authors.initials: N
authors.lastname: Miyata
authors.forename: Shu
authors.initials: S
authors.lastname: Hashimoto
authors.forename: Johan
authors.initials: J
authors.lastname: Van Lint
authors.forename: Chisei
authors.initials: C
authors.lastname: Ra
mesh_terms: D000818:Animals; D015153:Blotting, Western; D016207:Cytokines / Q000096:biosynthesis*; D004247:DNA / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors; D007202:Indicators and Reagents; D018793:Interleukin-13 / Q000096:biosynthesis / Q000235:genetics; D008407:Mast Cells / Q000187:drug effects / Q000378:metabolism*; D010766:Phosphorylation; D010957:Plasmids / Q000235:genetics; D011493:Protein Kinase C / Q000096:biosynthesis / Q000378:metabolism*; D051381:Rats; D017455:Receptors, IgE / Q000819:agonists*; D012190:Retroviridae / Q000235:genetics; D015398:Signal Transduction / Q000187:drug effects; D018808:Transcription Factor AP-1 / Q000378:metabolism*; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D016207:Cytokines; D004791:Enzyme Inhibitors; D007202:Indicators and Reagents; D018793:Interleukin-13; D017455:Receptors, IgE; D018808:Transcription Factor AP-1; D014409:Tumor Necrosis Factor-alpha; D004247:DNA; C089212:protein kinase D; D011493:Protein Kinase C