Upstream NFIL-6-like site located within a DNase I hypersensitivity region mediates LPS-induced transcription of the murine interleukin-1 beta gene.
Upstream NFIL-6-like site located within a DNase I hypersensitivity region mediates LPS-induced transcription of the murine interleukin-1 beta gene.
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DOI：
10.4049/jimmunol.153.1.143
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发表时间：
1994-07
影响因子：
4.4
通讯作者：
S. A. Godambe;David D. Chaplin;T. Takova;Clifford J. Bellone
中科院分区：
文献类型：
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作者：
S. A. Godambe;David D. Chaplin;T. Takova;Clifford J. Bellone
为了确定调节LPS刺激的IL-1 β基因转录的顺式作用元件，我们通过DNase I消化分析了小鼠IL-1 β基因。至少有两个过敏网站位于2200和2600 bp之间的单核吞噬细胞的转录起始位点的上游，但不是在IL-1 nonproducing未成熟的T细胞系。使用含有IL-1 β 5 '-侧翼区部分的报告构建体的转染，在DNase I超敏（DH）区内鉴定了LPS诱导IL-1 β基因表达所需的特异性DNA序列。两个特定的DNA序列的核因子结合的目标，使用电泳迁移率变动分析（EMSA）进行评估。一个位点含有NFIL-6结合的共有序列。该NFIL-6位点内的碱基取代导致LPS诱导的IL-1 β基因转录的实际消除。将NFIL-6样序列的多聚体直接引入同源或异源启动子的5'端，赋予了LPS诱导的转录，这表明该NFIL-6样共有位点是转录激活因子。抗-C/EBP β（NFIL-6）和抗-C/EBP δ（NFIL-6 β）Ab在NFIL-6样元件和单核细胞核提取物之间形成的复合物中鉴定了这两种蛋白质。C/EBP δ（NFIL-6 β）在使用来自IL-1非产生性T细胞系的提取物的复合物中未检测到。这些数据与内毒素激活鼠IL-1 β基因表达中对C/EBP β（NFIL-6）和C/EBP δ（NFIL-6 β）的要求一致。
To define the cis-acting elements that regulate LPS-stimulated IL-1 beta gene transcription, we analyzed the murine IL-1 beta gene by digestion with DNase I. At least two hypersensitive sites were located between 2200 and 2600 bp upstream of the transcription start site in mononuclear phagocytes, but not in an IL-1 nonproducing immature T cell line. Specific DNA sequences required for LPS induction of IL-1 beta gene expression were identified within the DNase I hypersensitive (DH) region using transfection of reporter constructs that contained portions of the IL-1 beta 5'-flanking region. Two specific DNA sequences were targets for nuclear factor binding as assessed with use of electrophoretic mobility shift analysis (EMSA). One site contained a consensus sequence for NFIL-6 binding. Base substitutions within this NFIL-6 site resulted in virtual elimination of LPS-induced IL-1 beta gene transcription. Introduction of multimers of the NFIL-6-like sequence immediately 5' to homologous or heterologous promoters conferred LPS-induced transcription, indicating that this NFIL-6-like consensus site was a transcriptional activator. Anti-C/EBP beta (NFIL-6) and anti-C/EBP delta (NFIL-6 beta) Abs identified both of these proteins in complexes formed between the NFIL-6-like element and mononuclear cell nuclear extracts. C/EBP delta (NFIL-6 beta) was not detected in complexes utilizing extracts from the IL-1 nonproducing T cell line. These data are consistent with the requirement for C/EBP beta (NFIL-6) and C/EBP delta (NFIL-6 beta) in the activation of murine IL-1 beta gene expression by endotoxin.