Interleukin-1 Activates an NF-B-like Factor in Osteoclast-like Cells

doi:10.1074/jbc.271.9.4605

Interleukin-1 Activates an NF-B-like Factor in Osteoclast-like Cells (*)

From the (1)Department of Biochemistry and
(2)Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-82 and the
(3)Department of Biochemistry, School of Dentistry, Showa University, 1-5-8, Hatanodai, Shinagawa-ku, Tokyo 142, Japan

^§ To whom correspondence should be addressed. Tel.: 81-92-641-1151 (ext. 5193); Fax: 81-92-641-8834.

Abstract

We investigated the NF-κB 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 1α,25-dihydroxyvitamin D and prostaglandin E 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-κB-like factor, which specifically bound to a κB motif DNA sequence, as detected by an electrophoretic mobility shift assay. IL-1 also activated NF-κB induction in osteoblasts. However, the NF-κB-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-κB 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-κB. Antibodies against c-Rel, Rel B, and p52 did not recognize the NF-κB-like factor. These results suggest that IL-1 activates an NF-κB-like factor in osteoclast-like cells, which contains p50 and p65-related proteins.

Footnotes

↵* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵¹ The abbreviations used are:

MNC

multinucleated cell(s)

TRAP

tartrate-resistant acid phosphatase

CT

calcitonin

NF-κB

nuclear factor κB

IL

interleukin

PG

prostaglandin

PMA

phorbol 12-myristate 13-acetate

PMSF

phenylmethanesulfonyl fluoride

EMSA

electrophoretic mobility shift assay

OCL

osteoclast-like cell(s)

POB

primary osteoblast(s)

DTT

dithiothreitol

MMP

matrix metalloproteinase

1α,25(OH) D

1α,25-dihydroxyvitamin D.
- Received July 17, 1995.
- Revision received January 2, 1996.