Harrington M A, Konicek B, Song A, Xia X L, Fredericks W J, Rauscher F J
Department of Medicine, Indiana University School of Medicine, Indianapolis 46202-5121.
J Biol Chem. 1993 Oct 5;268(28):21271-5.
Colony-stimulating factor-1 (CSF-1) is a member of the immediate early gene family, which is expressed in mitogen-stimulated quiescent fibroblasts. The biological effects of CSF-1 are multifaceted and include stimulation of the proliferation and differentiation of myeloid progenitors and activity of circulating monocytes and tissue-specific macrophages. Ablation of circulating levels of biologically active CSF-1 in mice leads to osteopetrosis and sterility, thus implicating a role for CSF-1 in bone remodeling and implantation. Identification of regulatory elements and cognate transcription factors that bind the csf-1 promoter and mediate such diverse expression patterns is of great interest. We identified a sequence element at -273 to -265 (relative to the transcription initiation site) in the murine csf-1 promoter, which contains overlapping consensus sequences for the Wilms' tumor protein (WT1), EGR-1, SP1, and SP3 proteins. WT1 and EGR-1 proteins produced in vitro bound to this sequence, and co-transfection of wt1 with a csf-1-cat reporter plasmid resulted in repression of promoter activity. Interestingly, nuclear extracts prepared from serum-stimulated C3H10T1/2 cells contained predominantly SP1 and SP3 binding activities, which recognized the -273 to -265 site. Thus repression of the csf-1 promoter by WT1 at this site may involve competition between SP1 family transcriptional activators and the WT1 repressor. Colony-stimulating factor-1 may be a physiologically relevant target gene for regulation by the WT1 transcription factor.
集落刺激因子-1(CSF-1)是即刻早期基因家族的成员,在有丝分裂原刺激的静止成纤维细胞中表达。CSF-1的生物学效应是多方面的,包括刺激髓系祖细胞的增殖和分化以及循环单核细胞和组织特异性巨噬细胞的活性。去除小鼠体内具有生物活性的CSF-1的循环水平会导致骨质石化和不育,因此表明CSF-1在骨重塑和着床中起作用。鉴定与csf-1启动子结合并介导这种多样表达模式的调控元件和相关转录因子具有重要意义。我们在小鼠csf-1启动子中鉴定出一个位于-273至-265(相对于转录起始位点)的序列元件,该元件包含与威尔姆斯瘤蛋白(WT1)、早期生长反应蛋白-1(EGR-1)、SP1和SP3蛋白重叠的共有序列。体外产生的WT1和EGR-1蛋白与该序列结合,并且将wt1与csf-1-cat报告质粒共转染导致启动子活性受到抑制。有趣的是,从血清刺激的C3H10T1/2细胞制备的核提取物主要含有SP1和SP3结合活性,它们识别-273至-265位点。因此,WT1在此位点对csf-1启动子的抑制可能涉及SP1家族转录激活因子与WT1阻遏物之间的竞争。集落刺激因子-1可能是WT1转录因子进行调控的生理相关靶基因。