Control of Prostate-Specific Gene Expression

前列腺特异性基因表达的控制

基本信息

批准号：
7859356
负责人：
ROBERT J. MATUSIK
金额：
$ 2.11万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-01 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7859356
关键词：
Adult Affinity Chromatography Androgen Receptor Androgen Response Element Androgens Base Pairing Binding Binding Proteins Binding Sites Biological Assay Cell Differentiation process Cell Line Cells Complex DNA DNase-I Footprinting Data Dependence Development Electrophoretic Mobility Shift Assay Elements Embryonic Development Endoderm Epithelial Epithelial Cells Epithelium Gene Expression Gene Transduction Agent Genes Goals JUN gene Knockout Mice LNCaP Link Liquid Chromatography Malignant neoplasm of prostate Maps Modeling Modification Molecular Weight Mus Nuclear Nuclear Protein Nuclear Proteins Pattern Peptides Play Positioning Attribute Prostate Prostate-Specific Antigen Prostatic Prostatic Diseases Prostatic hypertrophy Protein Isoforms Proteins Protocols documentation Publishing Receptor Signaling Regulation Reporter Genes Reporting Rodent Role Southwestern Blotting Spermine TS-1 Testing Tissues Transgenes Transgenic Animals Transgenic Mice Transgenic Organisms Work base c-myc Genes cancer cell cell determination forkhead protein human USF2 protein inhibitor/antagonist interest meetings novel probasin promoter public health relevance receptor binding research study response tandem mass spectrometry tool transcription factor transgene expression

项目摘要

DESCRIPTION (provided by applicant): Understanding the mechanism that controls prostate-specific gene expression has already resulted in the identification of FoxA1 as an important co-regulator of AR action as well as prostate development. We have now demonstrated that USF2 interacts with FoxA1 on multiple prostate-specific promoters. This indicates that USF2 functions to promote expression of genes associated with differentiation consistent with previously reported mechanism whereby USF2 inhibits proliferation by down regulating c-myc. Controlling prostate- specific gene expression with a complex of AR (signal dependent), FOXA1 (developmental cell specific for endoderm) and USF2 (differentiation specific) reveals a remarkable coming together of regulatory factors to dictate prostate-specific gene expression. Continuing work using tagged FOXA1 for affinity purification followed with Liquid Chromatography/Tandem Mass Spectrometry (LC-MS/MS) has identified 16 nuclear proteins that met the requirement of at least two peptide hits per protein in two separate experiments. Of these, 7 nuclear proteins can be either directly or indirectly linked to AR/FOXA1 action. Determining the factors that control prostate-specific gene expression has important implications for the understanding of cell fate during prostate development, androgen regulation of prostate disease, as well as understanding the fundamental cascade that controls cell determination and cell differentiation. Based upon published and our preliminary data, our Hypothesis is that by identifying the TFs that control prostate-specific gene expression, we are also identifying TF that play a critical role in prostate development. Three specific aims are proposed: Aim 1: To characterize the transcription factor complex of AR-regulated prostate-specific genes; Aim 2: To identify novel FOXA1 binding partners by tandem affinity purification followed with Liquid Chromatography/Tandem Mass Spectrometry (LC-MS/MS); Aim 3: To determine the function of identified TFs in prostate development and androgen dependence. Determining the remaining TFs that control prostate-specific gene expression has important implications for the understanding the factors that control androgen regulation of prostatic diseases such as hBPH and PCa. PUBLIC HEALTH RELEVANCE: Determining the factors that control androgen regulated prostate-specific gene expression has important implications for the understanding of cell fate during prostate development, androgen regulation of prostate disease, as well as understanding the fundamental cascade that controls cell determination and cell differentiation.

描述（由申请人提供）：了解控制前列腺特异性基因表达的机制已经导致将 FoxA1 鉴定为 AR 作用以及前列腺发育的重要共同调节因子。我们现在已经证明 USF2 与 FoxA1 在多个前列腺特异性启动子上相互作用。这表明USF2起到促进与分化相关的基因表达的作用，这与之前报道的USF2通过下调c-myc抑制增殖的机制一致。用 AR（信号依赖性）、FOXA1（内胚层特异性发育细胞）和 USF2（分化特异性）复合物控制前列腺特异性基因表达，揭示了调节因子的显着组合来决定前列腺特异性基因表达。使用标记的 FOXA1 进行亲和纯化，然后进行液相色谱/串联质谱 (LC-MS/MS) 的持续工作已鉴定出 16 个核蛋白，这些蛋白满足在两个单独的实验中每个蛋白至少有两个肽命中的要求。其中，7 个核蛋白可以直接或间接与 AR/FOXA1 作用相关。确定控制前列腺特异性基因表达的因素对于了解前列腺发育过程中的细胞命运、前列腺疾病的雄激素调节以及了解控制细胞决定和细胞分化的基本级联具有重要意义。根据已发表的数据和我们的初步数据，我们的假设是，通过识别控制前列腺特异性基因表达的 TF，我们还识别了在前列腺发育中发挥关键作用的 TF。提出了三个具体目标：目标 1：表征 AR 调节的前列腺特异性基因的转录因子复合物；目标 2：通过串联亲和纯化和液相色谱/串联质谱 (LC-MS/MS) 鉴定新型 FOXA1 结合伴侣；目标 3：确定已识别的转录因子在前列腺发育和雄激素依赖性中的功能。确定控制前列腺特异性基因表达的剩余转录因子对于了解控制 hBPH 和 PCa 等前列腺疾病雄激素调节的因素具有重要意义。公共健康相关性：确定控制雄激素调节的前列腺特异性基因表达的因素对于了解前列腺发育过程中的细胞命运、前列腺疾病的雄激素调节以及了解控制细胞决定和细胞分化的基本级联具有重要意义。