Characterization of the promoter for the human long pentraxin PTX3 - Role of NF-kappa B in tumor necrosis factor-alpha and interleukin-1 beta regulation
Characterization of the promoter for the human long pentraxin PTX3 - Role of NF-kappa B in tumor necrosis factor-alpha and interleukin-1 beta regulation
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DOI：
10.1074/jbc.272.13.8172
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发表时间：
1997-03-28
影响因子：
4.8
通讯作者：
Introna, M
中科院分区：
文献类型：
--
作者：
Basile, A;Sica, A;Introna, M
‘长五肽’是一个新的基因家族，它在其羧基末端保守了一个与典型的急性时相蛋白五肽(C-反应蛋白和血清淀粉样蛋白P组分)同源的五肽结构域，并获得了新的氨基端结构域。本文通过氯霉素乙酰转移酶和核糖核酸酶保护实验，研究了人‘’长五新‘基因PTX3的1371个核苷酸片段对8387人成纤维细胞中肿瘤坏死因子-α(TNF-α)和白介素1-β(IL-1β)的刺激作用。在同一细胞中，PTX3启动子对IL-6刺激没有反应。此外，在Hep 3B肝癌细胞系中未发现IL-1β和TNFα的反应性，最小启动子含有一个核因子-kappaB元件，该元件被证明是诱导所必需的，并且能够结合p50同源二聚体和p65异源二聚体，但不结合c-rel。该位点的突变体失去了结合核因子-kappaB蛋白的能力，并在功能检测中对肿瘤坏死因子α和白介素1β做出反应。位于核因子-kappa B位点附近的SP1和AP-1结合位点似乎并不对细胞因子的反应起主要作用。最后，与表达载体共转染实验证实，天然启动子含有一个功能性的核因子-kappaB位点。
The ''long pentraxins'' are an emerging family of genes that have conserved in their carboxy-terminal halves a pentraxin domain homologous to the prototypical acute phase protein pentraxins (C-reactive protein and serum amyloid P component) and acquired novel amino-terminal domains. In this report, a genomic fragment of 1371 nucleotides from the human ''long pentraxin'' gene PTX3 is characterized as a promoter on tumor necrosis factor-alpha (TNF alpha) and interleukin (IL)-1 beta exposure in transfected 8387 human fibroblasts by chloramphenicol acetyltransferase and RNase protection assays. In the same cells, the PTX3 promoter does not respond to IL-6 stimulation. Furthermore, IL-1 beta and TNF alpha responsiveness is not seen in the Hep 3B hepatoma cell line, The minimal promoter contains one NF-kappa B element which is shown to be necessary for induction and able to bind p50 homodimers and p65 heterodimers but not c-Rel. Mutants in this site lose the ability to bind NF-kappa B proteins and to respond to TNF alpha and IL-1 beta in functional assays. Sp1- and AP-1 binding sites lying in proximity to the NF-kappa B site do not seem to play a major role for cytokine responsiveness. Finally, cotransfection experiments with expression vectors validate that the natural promoter contains a functional NF-kappa B site.