The winged helix transcription factor HFH-4 is expressed during choroid plexus epithelial development in the mouse embryo
The winged helix transcription factor HFH-4 is expressed during choroid plexus epithelial development in the mouse embryo
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DOI：
10.1073/pnas.94.7.3094
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发表时间：
1997-04-01
影响因子：
11.1
通讯作者：
Costa, RH
中科院分区：
文献类型：
--
作者：
Lim, L;Zhou, HP;Costa, RH
哺乳动物肝细胞核因子 3 (HNF-3) 和果蝇同源基因叉头蛋白是细胞特异性转录因子广泛家族的原型，它们在翼状螺旋 DNA 结合域中具有同源性。这些哺乳动物家族成员之一，HNF-3/叉头同源物-4 (HFH-4)，是通过啮齿动物脑 cDNA 的 PCR 扩增分离出来的，并在成年细支气管上皮中表现出丰富的表达。在这项研究中，我们对特定阶段的小鼠胚胎进行了原位杂交，并报告了 HFH-4 基因在假定的和分化的脉络丛上皮中的新表达模式，脉络丛上皮负责脑脊液 (CSF) 蛋白的合成和分泌。我们还表明，HFH-4 在共转染测定中是一种有效的转录激活剂，并定义了几个对 HFH-4 转录活性重要的蛋白质序列。我们使用重组 HFH-4 蛋白进行体外 DNA 结合位点选择，并确定 HFH-4 蛋白识别 DNA 共有序列 HWDTGTTTGTTTA 或 KTTTGTTGTTKTW（其中 H 不是 G，W 是 A 或 T，D 不是 C，K 是 G 或 T）。我们使用此 HFH-4 共识来识别脉络丛上皮中潜在的 HFH-4 靶基因，并证明这些启动子序列在电泳迁移率变动测定中与重组 HFH-4 蛋白结合，重组 HFH-4 与人凝血酶原、β 淀粉样前体蛋白、α 1-抗胰凝乳蛋白酶、囊性纤维化跨膜电导调节剂的启动子区域形成特异性蛋白质-DNA 完成体以及啮齿动物 α2-巨球蛋白、生长激素受体和胰岛素样生长因子 II 基因。此外，我还确定了细支气管上皮中假定的 HFH-4 靶基因，包括克拉拉细胞分泌蛋白基因和 HNF-3 α 基因，HNF-3 α 基因是参与细支气管上皮基因转录调节的翼状螺旋家族成员。为了支持这些结合研究，共转染测定表明 HFH-4 增强了 INF-3 α 和克拉拉细胞分泌蛋白启动子区域的表达。
Mammalian hepatocyte nuclear factor-3 (HNF-3) and the Drosophila homeotic gene fork head proteins are prototypes of an extensive family of cell-specific transcription factors that share homology In the winged helix DNA-binding domain. One of these mammalian family members, HNF-3/fork head homolog-4 (HFH-4), was isolated by PCR amplification of rodent brain cDNA and exhibits abundant expression in the adult bronchiolar epithelium. In this study, we performed in situ hybridization of stage-specific mouse embryos and report on a novel expression pattern of the HFH-4 gene in both the presumptive and differentiated choroid plexus epithelium, which is responsible for the synthesis and secretion of cerebrospinal fluid (CSF) proteins. We also showed that HFH-4 is a potent transcriptional activator in cotransfection assays and defined several protein sequences important for HFH-4 transcriptional activity. We used in vitro DNA-binding site selection with recombinant HFH-4 protein and determined that the HFH-4 protein recognizes the DNA consensus sequences HWDTGTTTGTTTA or KTTTGTTGTTKTW (where H is not G, W is A or T, D is not C, and K is G or T). We used this HFH-4 consensus to identify potential HFH-4 target gents in the choroid plexus epithelium and demonstrated that these promoter sequences bind to recombinant HFH-4 protein in electrophoretic mobility shift assays, Recombinant HFH-4 formed specific protein-DNA completes with the promoter regions of the human prothrombin, beta amyloid precursor protein, alpha 1-antichymotrypsin, cystic fibrosis transmembrane conductance regulator and rodent alpha 2-macroglubulin, growth hormone receptors, and insulin-like growth factor II genes. Furthermore, me identified putative HFH-4 target genes in the bronchiolar epithelium including the clara cell secretory protein gene and the HNF-3 alpha gene, a winged helix family member involved in the transcriptional regulation of genes in the bronchiolar epithelium. In support of these binding studies, cotransfection assays show that HFH-4 potentiates expression of the INF-3 alpha and clara cell secretory protein promoter regions.