Washizu J, Nishimura H, Nakamura N, Nimura Y, Yoshikai Y
Laboratory of Host Defense, Research Institute for Disease Mechanism and Control, and First Department of Surgery, Nagoya University School of Medicine, 65 Tsurumai-cho Showa-ku, Nagoya 466, Japan.
Immunogenetics. 1998 Jun;48(1):1-7. doi: 10.1007/s002510050393.
We cloned the 5' upstream region of IL-15 genomic DNA and examined promoter activity in macrophages stimulated with lipopolysaccharide(LPS). The 1.2 kilobase (kb) fragment of the 5' upstream region contained binding elements for LPS-inducible transcription factors such as NFIL-6 or NF-kappaB. Determined by luciferase assay following transient transfection in the J774A.1 macrophage cell line, the 1.2 kb of the 5' upstream region exhibited high promoter activity in response to LPS, while promoter activity was significantly reduced by the 5' deletion of 313 base pairs containing the NF-kappaB binding motif. Nuclear protein prepared from LPS-stimulated macrophages formed a complex with the NF-kappaB binding sequence of the IL-15 promoter. Taken together, the binding of nuclear protein to the NF-kappaB binding site is required for transcriptional activation of the IL-15 gene in LPS-stimulated macrophages.
我们克隆了白细胞介素-15（IL-15）基因组DNA的5'上游区域，并检测了脂多糖（LPS）刺激的巨噬细胞中的启动子活性。5'上游区域的1.2千碱基（kb）片段包含LPS诱导转录因子如NFIL-6或核因子κB（NF-κB）的结合元件。通过在J774A.1巨噬细胞系中瞬时转染后的荧光素酶测定确定，5'上游区域的1.2 kb在响应LPS时表现出高启动子活性，而包含NF-κB结合基序的313个碱基对的5'缺失显著降低了启动子活性。从LPS刺激的巨噬细胞制备的核蛋白与IL-15启动子的NF-κB结合序列形成复合物。综上所述，核蛋白与NF-κB结合位点的结合是LPS刺激的巨噬细胞中IL-15基因转录激活所必需的。
