Functional analysis of the WT1-BASP1 transcriptional repressor complex

WT1-BASP1转录抑制复合物的功能分析

• 批准号: 8631091
• 负责人: KATHRYN MEDLER
• 金额: $ 29.37万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8631091
• 关键词: Adult; Apoptosis; Binding; Biology; Cell Line; Cells; Childhood; Clinical Trials; Complex; Data; Development; Disease; Embryo; Event; Gene Expression; Gene Targeting; Genes; Genetic Transcription; Goals; Growth; HDAC1 gene; Histone Deacetylase; Immunotherapy; Kidney; Light; Link; Malignant Childhood Neoplasm; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Modeling; Multiprotein Complexes; N-terminal; Nephroblastoma; Oncogenes; Oncogenic; Peptides; Phosphatidylinositol 4,5-Diphosphate; Phospholipids; Play; Process; Proteins; RNA Polymerase II; Regulation; Renal carcinoma; Role; Site; Solid; Transcription Coactivator; Transcription Repressor/Corepressor; Transcriptional Activation; Transcriptional Regulation; Tumor Suppressor Genes; Tumor Suppressor Proteins; WT1 Protein; WT1 gene; biological systems; cancer therapy; chromatin remodeling; cofactor; design; gene repression; insight; leukemia; malignant breast neoplasm; member; myristoylation; novel; podocyte; prohibitin; promoter; therapeutic target; transcription factor; tumor; tumorigenesis

DESCRIPTION (provided by applicant): The Wilms' tumor 1 protein is a developmental transcription factor that can either activate or repress genes involved in growth, apoptosis and differentiation. How these disparate activities of WT1 are regulated is not clear, but have important implications in determining its role in multiple cancers. Our long-term goal is to understand the mechanisms by which WT1 regulates transcription in development and disease. BASP1 is a critical regulator of WT1 that switches its function from a transcriptional activator to a repressor, and acts by binding to WT1 at the control regions of target genes. We have made the novel finding that the N-terminal myristoylation of BASP1 and the capacity of BASP1 to interact with phosphatidyinositol 4,5-bisphosphate (PIP2) is critical for its transcriptional cosuppressor function. Moreover, our recent data demonstrate that BASP1 is part of a large multiprotein complex that contains the transcriptional repressor prohibitin and histone deacetylase activity. The goal of the current proposal is to provide critical new insights into the mechanisms involved in the regulation of transcription by WT1-BASP1 and the novel role of PIP2. The specific aims are to; 1. Elucidate the mechanisms by which BASP1-PIP2 interactions control the transcription function of WT1. The role of PIP2 in transcriptional regulation by WT1-BASP1 through chromatin remodeling and RNA polymerase II activity will be investigated using ChIP, transcription analysis, and colocalization approaches. These studies will provide new insights into a novel mechanism of transcriptional repression by gene-specific regulators. 2. Isolate the components of the BASP1 complex and study their function in WT1 regulation. We have recently identified the transcriptional repressor prohibitin and histone deacetylase I as components of the BASP1 complex, which will be analyzed further to determine their mechanism of action. The other components of the complex will be identified and their role in BASP1-mediated transcriptional repression elucidated. 3. Determine the role of the BASP1 complex and PIP2 in the control of differentiation by WT1. A cell line model of podocyte differentiation that is dependent on the activities of WT1 and BASP1 will be employed to identify the critical components of the BASP1 complex, and uncover the role of PIP2 in WT1-dependent gene expression that drives the differentiation process.

描述（由申请人提供）： Wilms的肿瘤1蛋白是一种发育转录因子，可以激活或抑制与生长，凋亡和分化有关的基因。如何调节WT1的这些不同活动尚不清楚，但是在确定其在多种癌症中的作用方面具有重要意义。我们的长期目标是了解WT1调节发育和疾病转录的机制。 BASP1是WT1的关键调节因子，将其功能从转录激活因子转换为阻遏物，并通过在靶基因的控制区域与WT1结合起作用。我们已经提出了一个新的发现，即BASP1的N末端肉豆蔻酰化以及BASP1与磷脂酰辛醇4,5-双磷酸（PIP2）相互作用的能力对于其转录固定剂函数至关重要。此外，我们最近的数据表明，BASP1是包含转录抑制剂禁止素和组蛋白脱乙酰基酶活性的大型多蛋白络合物的一部分。 当前建议的目的是对WT1-BASP1调节转录和PIP2的新作用的机制提供关键的新见解。具体目标是； 1。阐明BASP1-PIP2相互作用控制WT1的转录函数的机制。 PIP2在WT1-BASP1通过染色质重塑和RNA聚合酶II活性中的转录调控中的作用将使用芯片，转录分析和共定位方法研究。这些研究将为基因特异性调节剂的转录抑制机制提供新的见解。 2。隔离BASP1复合物的成分，并研究其在WT1调节中的功能。我们最近将转录阻遏物禁止素和组蛋白脱乙酰基酶I确定为BASP1复合物的组成部分，将进一步分析以确定其作用机理。该复合物的其他组成部分将被识别，并在BASP1介导的转录抑制中阐明它们的作用。 3。确定BASP1复合物和PIP2在WT1分化的控制中的作用。将采用依赖于WT1和BASP1活性的足细胞分化的细胞系模型来识别BASP1复合物的关键成分，并揭示PIP2在WT1依赖性基因表达中的作用，从而驱动分化过程。