Interleukin-1 alpha activates an NF-kappaB-like factor in osteoclast-like cells.
We investigated the NF-kappaB transcription factor in osteoclast-like cells. Osteoclast-like cells were differentiated from mouse bone marrow cells in co-culture with mouse calvaria-derived primary osteoblasts in the presence of 1alpha,25-dihydroxyvitamin D3 and prostaglandin E2 in collagen gel-coated dishes. We enriched osteoclast-like cells from the co-cultures by Pronase treatment. When the enriched osteoclast-like cells were treated with phorbol 12-myristate 13-acetate, interleukin-1 (IL-1), calcitonin, or macrophage colony-stimulating factor, only IL-1 activated an NF-kappaB-like factor, which specifically bound to a kappaB motif DNA sequence, as detected by an electrophoretic mobility shift assay. IL-1 also activated NF-kappaB induction in osteoblasts. However, the NF-kappaB-like factor induced by IL-1-stimulated osteoclast-like cells is of smaller molecular size than the factor in osteoblasts, as shown by an electrophoretic mobility shift assay. The NF-kappaB activity of osteoclast-like cells was recognized completely by antibodies against the p50 subunit, and only partially by antibodies against the p65 subunit of NF-++kappaB. Antibodies against c-Rel, Rel B, and p52 did not recognize the NF-kappaB-like factor. These results suggest that IL-1 activates an NF-kappaB-like factor in osteoclast-like cells, which contains p50 and p65-related proteins.
Jimi
E
E
Department of Biochemistry, Faculty of Dentistry, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-82, Japan.
Ikebe
T
T
Takahashi
N
N
Hirata
M
M
Suda
T
T
Koga
T
T
D000818:Animals; D001483:Base Sequence; D001853:Bone Marrow / Q000378:metabolism; D001854:Bone Marrow Cells; D002116:Calcitonin / Q000494:pharmacology; D002117:Calcitriol / Q000494:pharmacology*; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism*; D018920:Coculture Techniques; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D015232:Dinoprostone / Q000494:pharmacology*; D005269:Femur; D007375:Interleukin-1 / Q000494:pharmacology*; D016173:Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism*; D015345:Oligonucleotide Probes; D010010:Osteoclasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D012886:Skull / Q000166:cytology / Q000502:physiology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013977:Tibia
D004268:DNA-Binding Proteins; D007375:Interleukin-1; D016328:NF-kappa B; D015345:Oligonucleotide Probes; D016173:Macrophage Colony-Stimulating Factor; D002116:Calcitonin; D004247:DNA; D002117:Calcitriol; D015232:Dinoprostone; D013755:Tetradecanoylphorbol Acetate