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（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FOXA1鉴定为AR作用的重要共同调节剂以及前列腺发育。现在，我们已经证明了USF2在多个前列腺特异性启动子上与FOXA1相互作用。这表明USF2的功能是促进与先前报道的机制一致的分化相关的基因表达，从而通过调节C-MYC来抑制增殖。用AR（信号依赖性），FOXA1（特异性内胚层的发育细胞）和USF2（分化特异性）来控制前列腺特异性基因表达，这揭示了调节因子的显着结合，以决定前列腺特异性基因表达。使用标记的FOXA1进行亲和力纯化，然后进行液相色谱/串联质谱法（LC-MS/MS）的持续工作，已经确定了16种核蛋白，这些核蛋白在两个单独的实验中满足了每个蛋白质至少两次肽的要求。其中，7个核蛋白可以直接或间接与AR/FOXA1作用联系起来。确定控制前列腺特异性基因表达的因素对在前列腺发育过程中对细胞命运的理解，前列腺疾病的雄激素调节以及了解控制细胞测定和细胞分化的基本级联反应具有重要意义。基于已发表和我们的初步数据，我们的假设是，通过识别控制前列腺特异性基因表达的TF，我们还确定了在前列腺发育中起关键作用的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：通过串联亲和力纯化确定新型的FOXA1结合伴侣，然后使用液相色谱/串联质谱（LC-MS/MS）来识别foxa1结合伙伴；目标3：确定已鉴定的TF在前列腺发育和雄激素依赖性中的功能。确定剩余的TF控制前列腺特异性基因表达对理解控制前列腺疾病（例如HBPH和PCA）的雄激素调节的因素具有重要意义。公共卫生相关性：确定控制雄激素调节前列腺特异性基因表达的因素对在前列腺发育过程中对细胞命运的理解，前列腺疾病的调节以及了解控制细胞确定和细胞分化的基本级联反应具有重要意义。