Inhibition of the production of rat cytokine-induced neutrophil chemoattractant (CINC)-1, a member of the interleukin-8 family, by adenovirus-mediated overexpression of IkappaBalpha.
Cytokine-induced neutrophil chemoattractant (CINC)-1, a counterpart of the human growth-regulated gene product (GRO) of the interleukin-8 family, has been suggested to play critical roles as a mediator of inflammatory reactions with neutrophil infiltration in rats. NF-kappaB has been speculated to be involved in the production of CINC-1, since the NF-kappaB-binding domain is important for the CINC-1 promoter activity in several of our reporter assays. In the present study, we examined the effects of an overexpression of IkappaBalpha, a specific natural inhibitor of NF-kappaB, on CINC-1 production. For this purpose, we constructed two recombinant adenoviruses, AxCAIkappaBalpha and AxCAmutantIkappaBalpha, which express respectively wild IkBa and a mutated nondegradable IkappaBalpha in which serine residues 32 and 36 are replaced by alanine residues. Transfecting wild and mutant IkBa by these adenovirus vectors inhibited NF-kappaB activation and CINC-1 production, which were both caused by IL-1beta stimulation in the normal rat kidney epithelial cell line NRK-52E. We also showed that the nondegradable mutant IkappaBalpha was approximately 30 times more potent than the wild type in inhibiting CINC-1 production. These findings demonstrate that CINC-1 production with NF-kappaB activation is primarily regulated by non-phosphorylated IkBa in the cytoplasm.
Takaishi
K
K
Institute of Cytosignal Research, Inc., Hiromachi, Shinagawa-ku, Tokyo 140-8710, Japan. takais@shina.sankyo.co.jp
Ohtsuka
T
T
Tsuneyoshi
S
S
Maehara
N
N
Harada
M
M
Yoshida
H
H
Watanabe
K
K
Tsurufuji
S
S
D000256:Adenoviridae / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D054360:Chemokine CXCL1; D019743:Chemokines, CXC; D002630:Chemotactic Factors / Q000494:pharmacology*; D003593:Cytoplasm / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000378:metabolism; D006131:Growth Inhibitors / Q000037:antagonists & inhibitors*; D006133:Growth Substances / Q000494:pharmacology*; D006801:Humans; D020672:I-kappa B Proteins; D036341:Intercellular Signaling Peptides and Proteins; D007375:Interleukin-1 / Q000494:pharmacology; D009154:Mutation; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism; D051381:Rats; D013997:Time Factors
C475829:CXCL1 protein, human; D054360:Chemokine CXCL1; D019743:Chemokines, CXC; D002630:Chemotactic Factors; C513297:Cxcl1 protein, rat; D004268:DNA-Binding Proteins; D006131:Growth Inhibitors; D006133:Growth Substances; D020672:I-kappa B Proteins; D036341:Intercellular Signaling Peptides and Proteins; D007375:Interleukin-1; D016328:NF-kappa B; C000606204:NFKBIA protein, human; C000606206:Nfkbia protein, rat; D000072000:NF-KappaB Inhibitor alpha