抄録
The JE/MCP-1 gene is an immediate-early gene, and its product is a CC chemokine that attracts monocytes, basophils and T lymphocytes. JE/MCP-1 gene expression is induced by various inflammatory stimuli, but its transcriptional mechanism is not fully understood. To address this question, we obtained two subclones from a parental RAW264.7 cell line, one subline with low JE/MCP-1-producing capacity (named RAW.c11) and the other with high JE/MCP-1-producing capacity (named RAW.c25), in response to lipopolysaccharide (LPS). These subclones have no significant differences in CD14 expression, nitric oxide production, or production of other cytokines, including TNF-α or 1α/lβ. In electrophoretic mobility shift assays (EMSA), there were no significant differences in DNA binding to the NF-κB-consensus sequence and interferon regulatory factor (IRF)-1,2 binding sequences. However, significantly higher binding activity to the NF-κB-like sequence (κB-3), which is located in the promoter region of the JE/MCP-1 gene, was shown by a high producer subclone than by a low producer subclone. Transient transfection analysis using deletion mutants of a 0.5-kb region from -467 to +59 identified an LPS-responsive region in a κB-3 site(from -169 to -132) in the high producer subclone. Mutation of this site markedly reduced sensitivity to LPS in the high producer subclone. These data suggest that a yet undefined nuclear factor may be involved in differential JE/MCP-1 gene transcription. (C) Academic Press.
| 元の言語 | English |
|---|---|
| ページ（範囲） | 207-219 |
| ページ数 | 13 |
| ジャーナル | Cytokine |
| 巻 | 12 |
| 発行部数 | 3 |
| DOI | |
| 出版物ステータス | Published - 2000 3 |
| 外部発表 | Yes |
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ASJC Scopus subject areas
- Endocrinology
- Molecular Biology
- Immunology
- Immunology and Allergy
これを引用
Differential induction of JE/MCP-1 in subclones from a murine macrophage cell line, raw 264.7 : Role of κB-3 binding protein. / Ueno, Masaya; Sonoda, Yoshiko; Tago, Megumi; Mukaida, Naofumi; Nose, Kiyoshi; Kasahara, Tadashi.
：: Cytokine, 巻 12, 番号 3, 03.2000, p. 207-219.研究成果: Article
}
TY - JOUR
T1 - Differential induction of JE/MCP-1 in subclones from a murine macrophage cell line, raw 264.7
T2 - Role of κB-3 binding protein
AU - Ueno, Masaya
AU - Sonoda, Yoshiko
AU - Tago, Megumi
AU - Mukaida, Naofumi
AU - Nose, Kiyoshi
AU - Kasahara, Tadashi
PY - 2000/3
Y1 - 2000/3
N2 - The JE/MCP-1 gene is an immediate-early gene, and its product is a CC chemokine that attracts monocytes, basophils and T lymphocytes. JE/MCP-1 gene expression is induced by various inflammatory stimuli, but its transcriptional mechanism is not fully understood. To address this question, we obtained two subclones from a parental RAW264.7 cell line, one subline with low JE/MCP-1-producing capacity (named RAW.c11) and the other with high JE/MCP-1-producing capacity (named RAW.c25), in response to lipopolysaccharide (LPS). These subclones have no significant differences in CD14 expression, nitric oxide production, or production of other cytokines, including TNF-α or 1α/lβ. In electrophoretic mobility shift assays (EMSA), there were no significant differences in DNA binding to the NF-κB-consensus sequence and interferon regulatory factor (IRF)-1,2 binding sequences. However, significantly higher binding activity to the NF-κB-like sequence (κB-3), which is located in the promoter region of the JE/MCP-1 gene, was shown by a high producer subclone than by a low producer subclone. Transient transfection analysis using deletion mutants of a 0.5-kb region from -467 to +59 identified an LPS-responsive region in a κB-3 site(from -169 to -132) in the high producer subclone. Mutation of this site markedly reduced sensitivity to LPS in the high producer subclone. These data suggest that a yet undefined nuclear factor may be involved in differential JE/MCP-1 gene transcription. (C) Academic Press.
AB - The JE/MCP-1 gene is an immediate-early gene, and its product is a CC chemokine that attracts monocytes, basophils and T lymphocytes. JE/MCP-1 gene expression is induced by various inflammatory stimuli, but its transcriptional mechanism is not fully understood. To address this question, we obtained two subclones from a parental RAW264.7 cell line, one subline with low JE/MCP-1-producing capacity (named RAW.c11) and the other with high JE/MCP-1-producing capacity (named RAW.c25), in response to lipopolysaccharide (LPS). These subclones have no significant differences in CD14 expression, nitric oxide production, or production of other cytokines, including TNF-α or 1α/lβ. In electrophoretic mobility shift assays (EMSA), there were no significant differences in DNA binding to the NF-κB-consensus sequence and interferon regulatory factor (IRF)-1,2 binding sequences. However, significantly higher binding activity to the NF-κB-like sequence (κB-3), which is located in the promoter region of the JE/MCP-1 gene, was shown by a high producer subclone than by a low producer subclone. Transient transfection analysis using deletion mutants of a 0.5-kb region from -467 to +59 identified an LPS-responsive region in a κB-3 site(from -169 to -132) in the high producer subclone. Mutation of this site markedly reduced sensitivity to LPS in the high producer subclone. These data suggest that a yet undefined nuclear factor may be involved in differential JE/MCP-1 gene transcription. (C) Academic Press.
KW - Chemokine
KW - IRF-2
KW - JE/MCP-1
KW - LPS
KW - NF-κB
UR - http://www.scopus.com/inward/record.url?scp=0033938394&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0033938394&partnerID=8YFLogxK
U2 - 10.1006/cyto.1999.0544
DO - 10.1006/cyto.1999.0544
M3 - Article
C2 - 10704247
AN - SCOPUS:0033938394
VL - 12
SP - 207
EP - 219
JO - Cytokine
JF - Cytokine
SN - 1043-4666
IS - 3
ER -