Identification of NF-kappaB in the marine fish Stenotomus chrysops and examination of its activation by aryl hydrocarbon receptor agonists.
Members of the Rel family of proteins have been identified in Drosophila, an echinoderm, Xenopus, birds and mammals. Dimers of Rel proteins form the transcription factor nuclear factor kappaB (NF-kappaB) that rapidly activates genes encoding cytokines, cell surface receptors, cell adhesion molecules and acute phase proteins. Evidence suggests that xenobiotic compounds also may alter the activation of NF-kappaB. This study had a dual objective of identifying members of the Rel family and examining their activation by xenobiotic compounds in a marine fish model, scup (Stenotomus chrysops). A DNA-protein crosslinking technique demonstrated that liver, kidney and heart each had at least three nuclear proteins that showed specific binding to an NF-kappaB consensus sequence, with molecular weights suggesting that the proteins potentially corresponded to mouse p50, p65 (RelA) and c-rel. In addition, an approximately 35kD NF-kappaB binding protein was evident in liver and kidney. The 50 kD protein was immunoprecipitated by mammalian p50-specific antibodies. The presence of Rel members in fish implied by those results was confirmed by RT-PCR cloning of a Rel homology domain (an apparent c-rel) from scup liver. NF-kappaB activation occurred in vehicle-treated fish, but this appeared to decrease over time. In fish treated with 0.01 or 1 mg 3,3',4,4', 5-pentachlorobiphenyl per kg, NF-kappaB activation in liver did not decrease, and there was a 6-8-fold increase in activation 16-18 days following treatment. Treatment with 10 mg benzo[a]pyrene/kg had no effect on NF-kappaB-DNA binding, either at 3 or 6 days following treatment. The data show that the Rel family of proteins is present in fish, represented at least by a p50/105 homologue, and support a hypothesis that some aryl hydrocarbon receptor agonists can activate NF-kappaB in vivo.
Schlezinger
J J
JJ
Biology Department, Redfield 342, MS 32, Woods Hole Oceanographic Institution, Woods Hole, MA 02543, USA.
Blickarz
C E
CE
Mann
K K
KK
Doerre
S
S
Stegeman
J J
JJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002645:Chickens; D016384:Consensus Sequence; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D005399:Fishes; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007668:Kidney / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D051379:Mice; D008862:Microsomes, Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism*; D010802:Phylogeny; D011078:Polychlorinated Biphenyls / Q000494:pharmacology*; D020569:Proto-Oncogene Proteins c-rel / Q000235:genetics* / Q000378:metabolism*; D018336:Receptors, Aryl Hydrocarbon / Q000819:agonists*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014422:Turkeys; D015262:Xenobiotics / Q000494:pharmacology; D014982:Xenopus laevis
D016328:NF-kappa B; D020569:Proto-Oncogene Proteins c-rel; D018336:Receptors, Aryl Hydrocarbon; D015262:Xenobiotics; D003577:Cytochrome P-450 Enzyme System; D011078:Polychlorinated Biphenyls