TY - JOUR
T1 - Functional characterization of human osteoclast inhibitory peptide-1 (OIP-1/hSca) gene promoter
AU - Srinivasan, Shanmugarajan
AU - Ito, Masahiro
AU - Kajiya, Hiroshi
AU - Key, L. Lyndon
AU - Johnson-Pais, Theresa L.
AU - Reddy, Sakamuri V.
N1 - Funding Information:
This work was supported by the National Institute of Health grants DE 12603 and Department of Defense Medical Research Award.
PY - 2006/4/12
Y1 - 2006/4/12
N2 - We have recently identified and characterized the human osteoclast (OCL) inhibitory peptide-1 (OIP-1/hSca), a member of Ly-6 gene family. OIP-1 is an important physiologic regulator of OCL development and bone resorption activity. To determine the molecular mechanisms that regulate OIP-1 gene expression in OCL precursor cells, we isolated and characterized the OIP-1/hSca gene (2 Kb) promoter sequence. IFN-γ (50 ng/ml) treatment of RAW 264.7 macrophage cells transfected with OIP-1 gene (- 1 to - 1988 bp relative to transcription start site) promoter-luciferase reporter plasmid demonstrated a significant (4 fold) increase in OIP-1 gene promoter activity. Sequence analysis of OIP-1 gene promoter region further identified a potential Stat-1 binding motif at - 1629 to - 1639 bp position. Stat-1 specific inhibitor, fludarabine (50 μM) abolished IFN-γ stimulated OIP-1 gene promoter activity. Electrophoretic mobility shift assay (EMSA) further confirmed activated Stat-1 binding to the OIP-1 gene promoter sequence suggesting that IFN-γ regulates OIP-1 gene expression in OCL precursor cells through a Stat-1 dependent signaling pathway. We further show that knock-down of TRADD enhances IFN-γ signaling to increase OIP-1 gene expression in OCL precursor cells. These results should provide insights into the molecular control of OIP-1 gene expression and inhibition of OCL activity in the bone microenvironment.
AB - We have recently identified and characterized the human osteoclast (OCL) inhibitory peptide-1 (OIP-1/hSca), a member of Ly-6 gene family. OIP-1 is an important physiologic regulator of OCL development and bone resorption activity. To determine the molecular mechanisms that regulate OIP-1 gene expression in OCL precursor cells, we isolated and characterized the OIP-1/hSca gene (2 Kb) promoter sequence. IFN-γ (50 ng/ml) treatment of RAW 264.7 macrophage cells transfected with OIP-1 gene (- 1 to - 1988 bp relative to transcription start site) promoter-luciferase reporter plasmid demonstrated a significant (4 fold) increase in OIP-1 gene promoter activity. Sequence analysis of OIP-1 gene promoter region further identified a potential Stat-1 binding motif at - 1629 to - 1639 bp position. Stat-1 specific inhibitor, fludarabine (50 μM) abolished IFN-γ stimulated OIP-1 gene promoter activity. Electrophoretic mobility shift assay (EMSA) further confirmed activated Stat-1 binding to the OIP-1 gene promoter sequence suggesting that IFN-γ regulates OIP-1 gene expression in OCL precursor cells through a Stat-1 dependent signaling pathway. We further show that knock-down of TRADD enhances IFN-γ signaling to increase OIP-1 gene expression in OCL precursor cells. These results should provide insights into the molecular control of OIP-1 gene expression and inhibition of OCL activity in the bone microenvironment.
KW - Interferon gamma
KW - Osteoclast inhibitory peptide-1 (OIP-1/hSca)
KW - Promoter
KW - Stat-1
KW - Transcription factor
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U2 - 10.1016/j.gene.2005.11.001
DO - 10.1016/j.gene.2005.11.001
M3 - Article
C2 - 16380218
AN - SCOPUS:33645212703
VL - 371
SP - 16
EP - 24
JO - Gene
JF - Gene
SN - 0378-1119
IS - 1
ER -