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.

项目总结 蛋白质ADP核糖化(ADPr)是一种动态的、依赖于NAD+的翻译后修饰。哺乳动物 多(ADP-核糖)聚合酶(PARP)蛋白催化ADPr靶向几种化学上不同的氨基酸 数百种底物蛋白质上的侧链官能团，介导多种正交信号 转导通路。增加这种复杂性的是形成ADP-核糖聚合物的可能性，这是一种过程 其中PARP1/2和TNKS1/2酶从单-ADPr位点拉长ADP-核糖链。突出显示 聚腺苷二磷酸核糖在生理和疾病中的重要性是：(I)PARP1/2的临床用途不断扩大 用于治疗DNA修复缺陷癌症的抑制剂，以及(Ii)TNKS1/2在Wnt/b-catenin信号转导和 包括切尔贝氏症在内的发育疾病的功能障碍。ADPr的异常活性也被报道为 心血管和神经生殖疾病的潜在原因，这些发现激发了强烈的 努力阐明PARP底物分布，确定PARP调节机制，并开发PARP 异构体特异性抑制物。然而，鉴于ADPr在蜂窝信令中的自由部署及其 拓扑复杂的化学性质，我们对特定的单和多ADPr位点如何影响的理解 蛋白质的功能和引发鲜明的生物活性却相对滞后。拟议的工作旨在填补这一点 通过开发新的方法来重建ADPr介导的信号事件来实现知识差距 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 丝氨酸ADPr到多肽和蛋白质上，完全控制修饰位点和ADP-核糖链的长度。 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。 核小体丝氨酸多聚ADPr的重塑和未知的特殊功能。我们现在正处于一个独特的 以我们的技术为基础，解决PARP生物学中的基本问题。我们将探索 控制多聚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中特定的生化输出。这样的信息可能会指导更多 有效的策略来识别和治疗依赖ADPr活性失调的疾病。

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