Control of Prostate-Specific Gene Expression

前列腺特异性基因表达的控制

• 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
• 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

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FoxA1被确定为AR作用和前列腺发育的重要共同调节因子。我们现在已经证明USF2在多个前列腺特异性启动子上与FoxA1相互作用。这表明USF2可以促进分化相关基因的表达，这与先前报道的USF2通过下调c-myc抑制增殖的机制一致。通过AR（信号依赖性）、FOXA1（内胚层发育细胞特异性）和USF2（分化特异性）的复合物控制前列腺特异性基因表达，揭示了一个显著的调控因子共同决定前列腺特异性基因表达。继续使用标记的FOXA1进行亲和纯化，然后使用液相色谱/串联质谱（LC-MS/MS）在两个单独的实验中鉴定出16个满足每个蛋白至少两个肽命中要求的核蛋白。其中，7种核蛋白可直接或间接与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理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理解控制细胞决定和细胞分化的基本级联具有重要意义。根据已发表的和我们的初步数据，我们的假设是，通过确定控制前列腺特异性基因表达的TF，我们也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；目的2：通过液相色谱/串联质谱（LC-MS/MS）串联亲和纯化鉴定新的FOXA1结合伙伴；目的3：确定已鉴定的tf在前列腺发育和雄激素依赖中的功能。确定控制前列腺特异性基因表达的剩余tf对于理解控制前列腺疾病（如hBPH和PCa）雄激素调节的因素具有重要意义。公共卫生相关性：确定雄激素调控的前列腺特异性基因表达的控制因素对于理解前列腺发育过程中的细胞命运、雄激素调控前列腺疾病以及理解控制细胞决定和细胞分化的基本级联具有重要意义。