Peterson R S, Clevidence D E, Ye H, Costa R H
Department of Biochemistry, University of Illinois at Chicago 60612-7334, USA.
Cell Growth Differ. 1997 Jan;8(1):69-82.
The hepatocyte nuclear factor-3 alpha (HNF-3 alpha) and -3 beta proteins share homology in the winged helix/fork head DNA binding domain and regulate cell-specific transcription in hepatocytes and respiratory epithelium. In this study, we used transfection assays to demonstrate that the -520 nucleotides upstream of the rat HNF-3 alpha gene were sufficient for cell-specific expression. We identified binding sites for a liver and kidney-enriched nuclear factor and a kidney-enriched protein that recognizes two distinct promoter elements. We showed that the rat HNF-3 alpha promoter binds the HNF-3 protein isoforms, which may serve an auto- and/or cross-regulatory role. Furthermore, we showed that cotransfection of the thyroid transcription factor-1 expression vector enhanced HNF-3 alpha promoter activity. We discuss these results with respect to the transcriptional induction of the HNF-3 alpha gene in respiratory epithelium during embryogenesis. Because the HNF-3 alpha promoter region bound nuclear factors in kidney extracts, we used in situ hybridization to demonstrate that it was expressed in the urothelium of the renal pelvis in adult and embryonic kidney. We also report on a novel expression pattern of HNF-3 alpha in the epithelium of the urinary bladder, penile urethra, and the prostate gland, and show that its expression in the intestinal epithelium increases from the proximal duodenum to distal ileum. We also demonstrate that HNF-3 alpha is abundantly expressed in the colonic epithelium. Furthermore, we use the HNF-3 DNA binding consensus sequence to identify putative target genes in the renal pelvis and gut epithelium.
肝细胞核因子-3α(HNF-3α)和-3β蛋白在翼状螺旋/叉头DNA结合结构域具有同源性,并调节肝细胞和呼吸道上皮细胞中的细胞特异性转录。在本研究中,我们通过转染实验证明大鼠HNF-3α基因上游-520个核苷酸足以实现细胞特异性表达。我们鉴定出了一种肝脏和肾脏富集核因子以及一种识别两个不同启动子元件的肾脏富集蛋白的结合位点。我们发现大鼠HNF-3α启动子能结合HNF-3蛋白异构体,这可能起到自我调节和/或交叉调节的作用。此外,我们还表明甲状腺转录因子-1表达载体的共转染增强了HNF-3α启动子活性。我们结合胚胎发育过程中呼吸道上皮中HNF-3α基因的转录诱导来讨论这些结果。由于HNF-3α启动子区域能结合肾脏提取物中的核因子,我们利用原位杂交证明其在成年和胚胎肾脏肾盂的尿路上皮中表达。我们还报道了HNF-3α在膀胱、阴茎尿道和前列腺上皮中的一种新表达模式,并表明其在肠上皮中的表达从十二指肠近端到回肠远端逐渐增加。我们还证明HNF-3α在结肠上皮中大量表达。此外,我们利用HNF-3 DNA结合共有序列来鉴定肾盂和肠道上皮中的潜在靶基因。