High affinity receptor for IgE stimulation activates protein kinase D augmenting activator protein-1 activity for cytokine producing in mast cells.
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.
Yamashita
Kyoko
K
Department of Molecular Cell Immunology and Allergology, Nihon University School of Medical Science, Tokyo, Japan.
Gon
Yasuhiro
Y
Shimokawa
Toshibumi
T
Nunomura
Satoshi
S
Endo
Daisuke
D
Miyata
Naoko
N
Hashimoto
Shu
S
Van Lint
Johan
J
Ra
Chisei
C
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
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