Regulation and function of site-specific protein poly-ADP-ribosylation

位点特异性蛋白质聚 ADP 核糖基化的调控和功能

• 批准号: 10668492
• 负责人: Glen Liszczak
• 金额: $ 41万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10668492
• 关键词: ADP ribosylation; Address; Adenosine Diphosphate Ribose; Amino Acids; Biochemical; Biological; Biology; Cardiovascular system; Cells; Chemicals; Cherubism; Clinical; Complex; DNA Damage; DNA Repair; DNA Repair Disorder; Development; Disease; Environment; Enzymes; Event; Functional disorder; Genetic Transcription; Knowledge; Length; Malignant Neoplasms; Mediating; Modification; Molecular; Nature; Nucleosomes; Output; Peptides; Physiology; Poly Adenosine Diphosphate Ribose; Poly(ADP-ribose) Polymerase Inhibitor; Poly(ADP-ribose) Polymerases; Polymers; Positioning Attribute; Post-Translational Protein Processing; Process; Protein Isoforms; Proteins; Regulation; Reporting; Research; Serine; Side; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Site; Site-Directed Mutagenesis; TNKS gene; Technology; Translations; Work; beta catenin; cancer therapy; chromatin remodeling; clinical application; developmental disease; human disease; inhibitor; new technology; novel strategies; programs; protein function; reconstitution

PROJECT SUMMARY Protein ADP-ribosylation (ADPr) is a dynamic, NAD+-dependent post-translational modification. The mammalian poly(ADP-ribose) polymerase (PARP) proteins that catalyze ADPr target several chemically distinct amino acid side chain functionalities on hundreds of substrate proteins to mediate a multitude of orthogonal signal transduction pathways. Adding to this complexity is the potential for ADP-ribose polymer formation, a process wherein the PARP1/2 and TNKS1/2 enzymes elongate ADP-ribose chains from mono-ADPr sites. Highlighting the importance of poly-ADP-ribose in physiology and disease are: (i) the expanding clinical utility of PARP1/2 inhibitors to treat DNA repair-deficient cancers, and (ii) TNKS1/2 function in Wnt/b-catenin signaling and dysfunction in developmental diseases including Cherubism. Aberrant ADPr activity has also been reported as an underlying cause of cardiovascular and neurogenerative diseases, and these findings have inspired intense efforts to elucidate PARP substrate profiles, determine PARP regulatory mechanisms, and develop PARP isoform-specific inhibitors. However, given the liberal deployment of ADPr in cellular signaling and its topologically complex chemical nature, our understanding of how specific mono- and poly-ADPr sites impact protein function and elicit distinct biological activities has lagged behind. The proposed work aims to fill this knowledge gap by developing novel approaches to reconstitute ADPr-mediated signaling events in highly controlled biochemical and cellular environments. We recently developed a chemoenzymatic strategy to install serine ADPr onto peptides and proteins with full control over modification site and ADP-ribose chain length. Using this technology, we identified critical molecular determinants of DNA damage-induced chromatin remodeling and uncovered specialized functions for nucleosome serine poly-ADPr. We are now in a unique position to build upon our technologies and address fundamental questions in PARP biology. We will explore mechanisms that govern poly-ADPr activity and investigate how different modification sites and accompanying polymer lengths encode for specific biochemical outputs throughout the cell. Such information may guide more effective strategies to identify and treat diseases that rely on dysfunctional ADPr activity.

项目概要 蛋白质 ADP-核糖基化 (ADPr) 是一种动态的、NAD+ 依赖性的翻译后修饰。哺乳动物 聚（ADP-核糖）聚合酶（PARP）蛋白催化 ADPr 靶标几种化学上不同的氨基酸 数百种底物蛋白上的侧链功能可介导大量正交信号 转导途径。 ADP-核糖聚合物形成的可能性增加了这种复杂性，这是一个过程 其中PARP1/2和TNKS1/2酶从单ADPr位点延长ADP-核糖链。突出显示 聚 ADP 核糖在生理学和疾病中的重要性是： (i) PARP1/2 不断扩大的临床应用 治疗 DNA 修复缺陷癌症的抑制剂，以及 (ii) TNKS1/2 在 Wnt/b-catenin 信号传导中发挥作用， 发育疾病中的功能障碍，包括天使主义。异常的 ADPr 活性也被报道为 心血管和神经系统疾病的根本原因，这些发现激发了人们的强烈兴趣 努力阐明 PARP 底物概况、确定 PARP 调控机制和开发 PARP 异构体特异性抑制剂。然而，鉴于 ADPr 在细胞信号传导中的自由部署及其 拓扑复杂的化学性质，我们对特定单和多 ADPr 位点如何影响的理解 蛋白质功能和引发独特的生物活性已经滞后。拟议的工作旨在填补这一空白 通过开发新方法来重建 ADPr 介导的信号事件的高度知识差距 受控的生化和细胞环境。我们最近开发了一种化学酶策略来安装 将丝氨酸 ADPr 附着到肽和蛋白质上，完全控制修饰位点和 ADP-核糖链长度。 利用这项技术，我们确定了 DNA 损伤诱导染色质的关键分子决定因素 重塑并发现核小体丝氨酸聚 ADPr 的特殊功能。我们现在正处于一个独特的 建立我们的技术并解决 PARP 生物学中的基本问题。我们将探索 控制聚 ADPr 活性的机制并研究不同的修饰位点和伴随的 聚合物长度编码整个细胞的特定生化输出。这些信息可能会指导更多 识别和治疗依赖 ADPr 活性功能失调的疾病的有效策略。

项目成果

A Protein Semisynthesis-Based Strategy to Investigate the Functional Impact of Linker Histone Serine ADP-Ribosylation.

• DOI: 10.1021/acschembio.2c00091
• 发表时间: 2022-04-15
• 期刊: ACS CHEMICAL BIOLOGY
• 影响因子: 4
• 作者: Tashiro, Kyuto;Mohapatra, Jugal;Brautigam, Chad A.;Liszczak, Glen
• 通讯作者: Liszczak, Glen

Asymmetric nucleosome PARylation at DNA breaks mediates directional nucleosome sliding by ALC1.

• DOI: 10.1038/s41467-024-45237-8
• 发表时间: 2024-02-02
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Bacic, Luka;Gaullier, Guillaume;Mohapatra, Jugal;Mao, Guanzhong;Brackmann, Klaus;Panfilov, Mikhail;Liszczak, Glen;Sabantsev, Anton;Deindl, Sebastian
• 通讯作者: Deindl, Sebastian

Chemoenzymatic and Synthetic Approaches To Investigate Aspartate- and Glutamate-ADP-Ribosylation.

研究天冬氨酸和谷氨酸-ADP-核糖基化的化学酶法和合成方法。

• DOI: 10.1021/jacs.3c03771
• 发表时间: 2023
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Tashiro K