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J Biol Chem, Vol. 275, Issue 12, 8331-8340, March 24, 2000
From the Murine osteoclast precursors and osteoblasts
express the integrin This paper is dedicated to our late secretary, Jane Wodicker.
Sp1/Sp3 and PU.1 Differentially Regulate
5
Integrin Gene Expression in Macrophages and Osteoblasts*
,,,¶
Department of Pathology and
§ Division of Bone and Mineral Diseases, Washington
University School of Medicine, St. Louis, Missouri 63110
v
5, the
appearance of which on the cell surface is controlled by the
5, and not the v, subunit. Here, we show
that a 173-base pair proximal region of the 5 promoter
mediates 5 basal transcription in macrophage (osteoclast
precursor)-like and osteoblast-like cells. DNase I footprinting reveal
four regions (FP1-FP4) within the 173-base pair region, protected by
macrophage nuclear extracts. In contrast, osteoblast nuclear extracts
protect only FP1, FP2, and FP3. FP1, FP2, and FP3 bind Sp1 and Sp3 from
both macrophage and osteoblast nuclear extracts. FP4 does not bind
osteoblast proteins but binds PU.1 from macrophages. Transfection
studies show that FP1 and FP2 Sp1/Sp3 sites act as enhancers in both
MC3T3-E1 (osteoblast-like) and J774 (macrophage-like) cell lines,
whereas the FP3 Sp1/Sp3 site serves as a silencer. Mutation of the FP2 Sp1/Sp3 site totally abolishes promoter activity in J774 cells, with
only partial reduction in MC3T3-E1 cells. Finally, we demonstrate that
PU.1 acts as a 5 silencer in J774 cells but plays no
role in MC3T3-E1 cells. Thus, three Sp1/Sp3 sites regulate
5 gene expression in macrophages and osteoblast-like
cells, with each element exhibiting cell-type and/or
activation-suppression specificity.
*
This study was partially supported by National Institutes of
Health Grants AR42404 (to F. P. R.), DE05413 and AR32788 (to S. L. T.), and AR07033 and AR32087 (to L. V. A.); a grant from Shriners Hospital (to S. L. T.); and National Institutes of Health Individual National Research Service Award AR085020 (to X. F.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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