Binding of two nuclear complexes to a novel regulatory element within the human S100A9 promoter drives the. S100A9 gene expression
Binding of two nuclear complexes to a novel regulatory element within the human S100A9 promoter drives the. S100A9 gene expression
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DOI：
10.1074/jbc.m207990200
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发表时间：
2002-11-01
影响因子：
4.8
通讯作者：
Klempt, M
中科院分区：
文献类型：
--
作者：
Kerkhoff, C;Hofmann, HA;Klempt, M
S100A9也被称为MRP14，是一种钙结合蛋白，其表达在髓细胞分化过程中受到严格调控。本研究旨在研究S100A9基因的细胞类型和分化特异性转录调控。对MonoMac-6细胞中S100A9启动子的分析显示，在-400至- 374bp位置存在一个新的调控区域，称为髓系相关蛋白调控元件（MRE）。MRE缺失导致启动子活性降低5.2倍。通过电泳迁移率转移分析，鉴定出两个与该区域结合的核复合物，分别为mre结合复合物A （MbcA）和mre结合复合物B （MbcB）。通过诱变，mre结合基序可以缩小到一个12bp的区域。通过观察MRE结合复合体A或MRE结合复合体B的形成与S100A9基因表达以细胞类型特异性、激活和分化依赖的方式强烈相关，推断出MRE的相关性。此外，DNA亲和层析和Western blot研究表明，kruppel相关锌指蛋白和转录中介因子1 β (tif1 β)参与mre结合复合体，从而调节S100A9基因的表达。
S100A9, also referred to as MRP14, is a calcium-binding protein whose expression is tightly regulated during differentiation of myeloid cells. The present study was performed to study the cell type- and differentiation-specific transcriptional regulation of the S100A9 gene. Analysis of the S100A9 promoter in MonoMac-6 cells revealed evidence for a novel regulatory region from position -400 to -374 bp, termed myeloid-related protein regulatory element (MRE). MRE deletion resulted in a 5.2-fold reduction of promoter activity. By electrophoretic mobility shift analysis two nuclear complexes binding to this region were identified and referred to as MRE-binding complex A (MbcA) and MRE-binding complex B (MbcB). By mutagenesis the MRE-binding motif could be narrowed to a 12-bp region. The relevance of MRE is deduced from the observations that the formation of either MRE-binding complex A or MRE-binding complex B strongly correlated with S100A9 gene expression in a cell type-specific, activation- and differentiation-dependent manner. Moreover, DNA affinity chromatography and Western blot studies indicate that a Kruppel-related zinc finger protein and the transcriptional intermediary factor 1beta (TIF1beta) are involved in an MRE-binding complex, thereby regulating the S100A9 gene expression.