Molecular mechanisms regulating Poly(ADP-ribosyl)ation pathway in living cells

活细胞中多聚（ADP-核糖基）化途径的分子机制调节

• 批准号: 2231403
• 负责人: Alexei Tulin
• 金额: $ 74.07万
• 依托单位: University of North Dakota Main Campus
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2231403&HistoricalAwards=false
• 关键词: Molecular; mechanisms; regulating; Poly; ADP

Developmental processes are tightly regulated, and their disruption leads to pathologies. Chromatin factors regulate developmental processes by controlling the chromatin state of specific loci to coordinate expression of multiple developmental genes. Much remains to be learned about how chromatin is programmed and how active or inactive domains are maintained during development. Poly(ADP-ribose) polymerase 1 (PARP1) and poly(ADP-ribose) glycohydrolase (PARG) (PAR enzymes) play a vital role during organismal development by controlling chromatin state and transcription. In this project, researchers will use the PAR pathway as a model to understand how cells undergo quick, localized and reversible chromatin reprogramming to fine tune gene activity. The findings will enhance our knowledge of developmental gene regulation and facilitate new methodology for reprogramming genes, for example through activation of PAR enzymes present in repressed chromatin loci. This project will reinforce the research environment at University of North Dakota by providing local high school and undergraduate students of different backgrounds with new opportunities to learn fundamental and biomedical aspects of developmental processes.This project will focus on the study of molecular mechanisms of PARG enzyme regulation, using Drosophila as a model. The research will be driven by the hypothesis that regulation of PARG activity is crucial for tissue-specific and cell cycle-specific differences in poly(ADP-ribosyl)ation rates. The preliminary data show that PARG, which does not have a DNA-binding motif, has two distinct roles in chromatin: 1) PARP1-independent binding to gene body of silent genes, likely to repress their expression; 2) binding of PARP1-occupied active loci, likely to maintain their expression in cooperation with PARP1. This project will focus on characterizing the molecular mechanism targeting and regulating PAR enzymes in living cells in steady-state conditions and in mutant backgrounds. The experiments will probe 1) what mechanisms recruit PAR enzymes to specific loci and 2) what triggers and fine-tunes PAR-dependent processes, including underlying mechanisms. The outcomes will advance understanding of the PAR pathway and how it might be modulated in new ways.This project is jointly funded by the Genetic Mechanisms program in the Molecular and Cellular Biosciences Division of the Directorate for Biological Sciences and the Established Program to Stimulate Competitive Research (EPSCoR).This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

发育过程受到严格的调节，它们的破坏导致病理。 染色质因子通过控制特定基因座的染色质状态来协调多个发育基因的表达来调节发育过程。 关于如何对染色质进行编程以及在开发过程中如何保持活性或无活性域，还有许多待了解。 通过控制染色质状态和转录，聚（ADP-核糖）聚合酶1（PARP1）和聚（ADP-核糖）Glydrollase（PARG）（PAR酶）在有机体发育过程中起着至关重要的作用。 在该项目中，研究人员将使用PAR途径作为模型来了解细胞如何进行快速，局部和可逆的染色质重编程以微调基因活性。 这些发现将增强我们对发展基因调控的了解，并促进重编程基因的新方法，例如通过激活抑制染色质基因座中存在的PAR酶。 该项目将通过为当地的高中和不同背景的本科生提供新的机会来学习发展过程的基本和生物医学方面，从而增强北达科他大学的研究环境。该项目将专注于使用Drosophila作为模型的Parg酶调节的分子机制研究。 这项研究将由以下假设驱动：PARG活性的调节对于聚（ADP-核糖基）率的组织特异性和细胞周期特异性差异至关重要。 初步数据表明，没有DNA结合基序的PARG在染色质中具有两个不同的作用：1）与静音基因的基因体无关的结合，可能会抑制其表达。 2）PARP1占主动性基因座的结合，可能与PARP1合作保持表达。 该项目将着重于表征在稳态条件和突变背景下活细胞中靶向和调节PAR酶的分子机制。 实验将探测1）哪些机制将par酶募集到特定的基因座和2）触发哪些触发和微型核心依赖性过程，包括潜在的机制。 结果将提高人们对PAR通路的了解以及如何以新的方式进行调节。该项目由生物科学局的分子和细胞生物科学局的遗传机制计划共同资助，以及启发竞争性研究（EPSCOR）的既定范围的Intortrial Internation the Intortial Mertul of Supportial Worth的竞争性研究，是由NSF的Infortial Induromed Interial Inderial te Elem deem deem deem deem deem deem tedy aperial of deem，审查标准。