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NF‐κB elements contribute to junB inducibility by lipopolysaccharide in the murine macrophage cell line RAW264.7
Author(s) -
Frazier-Jessen Michelle R,
Thompson Cynthia D,
Brown Robert,
Rawat Rashmi,
Nordan Richard P,
Feldman Gerald M
Publication year - 2002
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(02)02295-0
Subject(s) - junb , microbiology and biotechnology , reporter gene , lipopolysaccharide , transcription factor , nfkb1 , electrophoretic mobility shift assay , cell culture , gene expression , biology , chemistry , gene , immunology , biochemistry , genetics
Macrophages respond to bacterial lipopolysaccharide (LPS) by activating latent cis ‐acting factors that initiate transcription of immediate early genes. One such immediate early gene, junB , is induced by LPS in macrophages within 30 min. To identify elements that mediate the induction of junB by LPS, upstream and downstream sequences flanking the junB gene were examined by transient expression in the RAW264.7 murine macrophage cell line using a luciferase reporter gene vector containing the junB minimal promoter. A >10‐fold enhancement was associated with a 222 bp region downstream of the junB promoter in response to LPS. Transient reporter assays demonstrated that multiple nuclear factor (NF) κB sites are required for inducibility of junB by LPS in RAW264.7 cells. Electrophoretic mobility shift assays confirmed binding of LPS‐induced nuclear proteins included p50/p65 heterodimers at these NF‐κB sites.