Regulation of Kidney-Specific Gene Expression

肾脏特异性基因表达的调控

• 批准号: 6517196
• 负责人: Peter Igarashi
• 金额: $ 28.08万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-04-15 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6517196
• 关键词: binding proteins; cadherins; cell differentiation; epithelium; gel mobility shift assay; gene expression; genetic library; genetic mapping; genetic promoter element; genetic regulation; genetic regulatory element; genetically modified animals; histogenesis; laboratory mouse; molecular cloning; polycystic kidney; protein binding; renal tubular transport; tissue /cell culture; transcription factor; transfection

DESCRIPTION (provided by applicant): The long-term objectives of this project are to elucidate the molecular mechanisms of kidney-specific gene expression and renal epithelial cell differentiation, and to understand how perturbations of these mechanisms cause abnormal kidney development in humans. In the previous project period, we cloned the promoter of a novel kidney-specific cadherin gene (Ksp-cadherin) and showed that a short region of the promoter correctly recapitulates the complete expression pattern of the gene in transgenic mice. The Ksp-cadherin promoter is active exclusively in tubular epithelial cells in the kidney and the developing genitourinary tract. In addition, the kidney-enriched transcription factors, HNF-l and HNF-3, were shown to bind to the promoter and stimulate transcriptional activity. The proposed studies will test the hypothesis that transcription factors that regulate tissue-specific expression of Ksp-cadherin play important roles in kidney-specific gene expression and renal epithelial cell differentiation. In the first specific aim, we wil develop a general method for kidney-specific gene targeting that will be used to inactivate the hepatocyte nuclear factor 113 gene (HNF-1b) in transgenic mice. Mutations of the human HNF-1b gene cause maturity-onset diabetes of the young (MODY) and renal developmental abnormalities. The proposed studies will test whether I-INF-1 b is required for renal tubulogenesis and kidney-specific gene expression, and will examine the effects of a dominant gain-of-function mutation of HNF- b on kidney development. The second specific aim is to identify and characterize additional transcription factors that regulate kidney-specific gene expression. The region of the Ksp-cadherin promoter that is required for tissue-specific expression will be narrowed down, and renal nuclear proteins that bind to the region will be identified. Binding proteins that are co-expressed with Ksp-cadherin in renal tubular epithelial cells will be characterized further. Taken together, these studies will provide new insights into the genetic regulation of renal epithelial cell differentiation and the pathogenesis of kidney abnormalities in MODY. Moreover, these studies will generate unique reagents that will be widely applicable to studies involving cell lineage analysis in the kidney and kidney-specific gene targeting.

描述（由申请人提供）：该项目的长期目标 旨在阐明肾脏特异性基因表达的分子机制 和肾上皮细胞分化，并了解扰动如何 这些机制导致人类肾脏发育异常。在 在之前的项目期间，我们克隆了一种新型肾脏特异性的启动子 钙粘蛋白基因（Ksp-cadherin）并显示启动子的一个短区域 正确地概括了该基因的完整表达模式 转基因小鼠。 Ksp-钙粘蛋白启动子仅在肾小管中活跃 肾脏和发育中的泌尿生殖道的上皮细胞。在 此外，肾脏富集的转录因子 HNF-1 和 HNF-3 显示与启动子结合并刺激转录活性。这 拟议的研究将检验以下假设：转录因子 调节 Ksp-cadherin 的组织特异性表达在 肾脏特异性基因表达和肾上皮细胞分化。在 第一个具体目标是，我们将开发一种针对肾脏特异性的通用方法 用于灭活肝细胞核因子的基因靶向 转基因小鼠中的 113 基因（HNF-1b）。人类HNF-1b基因突变导致 青少年发病的成年期糖尿病 (MODY) 与肾脏发育 异常。拟议的研究将测试 I-INF-1 b 是否是必需的 肾小管发生和肾脏特异性基因表达，并将检查 HNF-b显性功能获得性突变对肾脏的影响 发展。第二个具体目标是识别和表征其他 调节肾脏特异性基因表达的转录因子。该地区 组织特异性表达所需的 Ksp-钙粘蛋白启动子 将缩小范围，与该区域结合的肾核蛋白将 被识别。与 Ksp-钙粘蛋白共表达的结合蛋白 将进一步表征肾小管上皮细胞。综合起来， 这些研究将为肾脏的基因调控提供新的见解。 上皮细胞分化和肾脏异常的发病机制 么。此外，这些研究将产生独特的试剂，将被广泛应用 适用于涉及肾脏细胞谱系分析的研究 肾脏特异性基因靶向。