The Role of PARP-1 in Hormone-Regulated Transcription

PARP-1 在激素调节转录中的作用

• 批准号: 6854740
• 负责人: WILLIAM Lee KRAUS
• 金额: $ 23.7万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6854740
• 关键词: biological signal transduction; cell free system; cell line; chromatin; chromatin immunoprecipitation; clinical research; enzyme activity; enzyme mechanism; genetic regulation; genetic transcription; hormone regulation /control mechanism; immunofluorescence technique; metabolism; microarray technology; nicotinamide adenine dinucleotide; nuclear receptors; pentosyltransferase

DESCRIPTION (provided by applicant): The coordinated regulation of gene expression in response to hormonal signals is a fundamental process in biology. Lipophilic hormones (e.g., estrogens in vertebrates, and ecdysteroids in insects) comprise a structurally diverse array of compounds that play important roles in many physiological processes, including growth, development, reproduction, and metabolism. Aberrations in the signaling pathways controlled by these hormones lead to disease states, including developmental abnormalities and cancers. The lipophilic hormones exert their effects through nuclear receptor (NR) proteins that function as DNA-binding transcription factors in the chromatin environment of the nucleus. The long term objective of these studies is to test the broad hypothesis that transcriptional outcomes in NR signaling pathways are determined by physical and functional interactions among the receptors, their associated coregulatory proteins, and chromatin. In particular, we will investigate a possible link between metabolic cofactors and signal-regulated transcription, focusing on the role of poly (ADP-ribose) polymerase- 1 (PARP-1) in NR-dependent gene regulation. PARP-1 is an intriguing coregulatory protein whose activity is regulated by the metabolic cofactor nicotinamide adenine dinucleotide (NAD¿). We will use a variety of complementary experimental systems (e.g., human cells and Drosophila larvae) and approaches (e.g., biochemical, cell-based, and organismal) to examine the mechanisms underlying PARP-1 coregulatory activity with NRs. The approaches include a unique biochemical ("cell-free") assay that accurately recreates hormone-dependent transcription with chromatin templates assembled in vitro, as well as microarray-based chromatin immunoprecipitation assays ("CHIP on chips") and immunofluorescent staining of Drosophila polytene chromosomes to generate a global view of factor distribution at hormone-regulated genes. These studies should lead to significant advances in the way we analyze hormone-regulated gene expression and increase our understanding of how these processes occur in normal and disease states.

描述（由适用提供）：响应激素信号的基因表达的协调调节是生物学的基本过程。亲脂术（例如，脊椎动物中的雌激素和昆虫中的ecdy骨）包含结构上多样的化合物，这些化合物在许多物理过程中起着重要作用，包括生长，发育，繁殖和代谢。这些激素控制的信号传导途径中的像差导致疾病状态，包括发展异常和癌症。亲脂性骑术通过核受体（NR）蛋白发挥作用，这些蛋白在核素的染色质环境中充当DNA结合转录因子。 这些研究的长期目标是检验广泛的假设，即NR信号通路中的转录结果取决于接收器之间的物理和功能相互作用，其相关的核调节蛋白和染色质。特别是，我们将研究代谢辅助因子与信号调节的转录之间的可能联系，重点是聚（ADP-核糖）聚合酶-1（PARP-1）在NR依赖性基因调节中的作用。 PARP-1是一种有趣的核心调节蛋白，其活性受代谢辅助剂烟酰胺腺苷二核苷酸（NAD®）调节。我们将使用多种互补的实验系统（例如人类细胞和果蝇幼虫）和方法（例如，生化，基于细胞和有机物）的方法来检查使用NRS的PARP-1分支活性的机制。 The approaches include a unique biochemical ("cell-free") assay That accurately recreates horsene-dependent transcription with chromatin templates assembled in vitro, as well as microarray-based chromatin immunoprecipitation assays ("CHIP on chips") and immunofluorescent staining of Drosophila polytene chromosomes to generate a global view of factor distribution at horsene-regulated genes.这些研究应在分析骑马调节基因表达的方式上取得重大进展，并增加对这些过程在正常状态和疾病状态中如何发生的理解。