Structural Biochemistry of PARP-1

PARP-1 的结构生物化学

基本信息

批准号：
8583327
负责人：
John M Pascal
金额：
$ 38.34万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-12-01 至 2015-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8583327
关键词：
Adenosine Diphosphate Ribose Affinity Amino Acids Apoptotic Architecture BRCT Domain Binding Biochemical Biological Assay Biological Process Biology Catalytic Domain Cell Culture Techniques Cell Death Cell Death Signaling Process Cell Survival Cell physiology Cells Chromatin Chromosomal Rearrangement Clinical Trials Communication Complex Couples Crystallography DNA DNA Binding DNA Binding Domain DNA Damage DNA Repair DNA Repair Enzymes DNA Structure Development Enzymes Event Gene Expression Regulation Gene Mutation Genome Histones Inflammation Knowledge Malignant Neoplasms Mediating Modeling Modification Molecular Molecular Conformation Monitor Mutagenesis Organism Pathway interactions Personal Satisfaction Poly Adenosine Diphosphate Ribose Poly(ADP-ribose) Polymerases Polymers Post-Translational Protein Processing Proteins Regulation Research Risk Role Signal Transduction Stress Structural Biochemistry Structure Testing Therapeutic Therapeutic Agents Transcriptional Regulation Work Zinc Fingers base cancer therapy improved inhibitor/antagonist insight interest member next generation novel strategies programs public health relevance repaired response therapeutic target transcription factor

项目摘要

DESCRIPTION (provided by applicant): Breaks in the structure of DNA are a persistent stress on the integrity of the genome, and they pose a substantial risk of chromosomal rearrangement and genetic mutation that can challenge the well-being of an organism and promote the development of cancer. There are several cellular mechanisms that monitor the state of the genome and rapidly initiate repair mechanisms in response to DNA damage so that a healthy genome is passed on to the next generation. Poly(ADP-ribose) Polymerase-1, or PARP-1, is a primary responder to breaks in the structure of DNA. PARP-1 has a unique catalytic activity that synthesizes polymers of ADP-ribose as a posttranslational modification on target proteins, primarily on PARP-1 itself (automodification). Upon binding to DNA breaks, PARP-1 activity is "turned on" to modulate DNA damage repair pathways and thereby promote cell survival. In contrast, excessive DNA damage leads to an elevated level of PARP-1 activity that results in cell death. Regulation of PARP-1 activity is therefore a critical factor in determining the fate of a cell. Furthermore, inhibitors of PARP-1 have recently emerged as promising therapeutic agents for the treatment of cancer and inflammation. Despite a growing interest in PARP-1 inhibitors and the discovery of expanded roles for PARP-1 activity in DNA repair, transcriptional regulation, and apoptotic signaling, there are few insights into the mechanism of PARP-1 activity and regulation. The long-term objective of this research program is to establish at the molecular level the mechanisms that control PARP-1 activity. DNA damage is the most potent activator of PARP-1; therefore we have chosen to first focus on the mechanism of DNA-dependent activation of PARP-1. Using a combination of x-ray crystallography and biochemical analysis, the proposed research will advance our understanding of PARP-1 recognition of DNA damage, and PARP-1 interaction with chromatin (Specific Aim 1). These studies will provide the first views of PARP-1 bound to DNA and will therefore provide mechanistic insights that will significantly advance the PARP field of research, as well as having a more broad impact on the field of DNA repair and chromatin biology. The proposed work will demonstrate how multiple domains of PARP-1 collaborate to couple poly(ADP-ribose) synthesis to structure-specific DNA binding (Specific Aim 2). The detailed structural analysis of PARP-1 activity and regulation will improve current models of PARP-1 biological functions and potentially reveal novel strategies for specifically inhibiting PARP-1 activity.

描述（由申请人提供）：DNA结构中断是对基因组完整性的持续压力，它们构成了染色体重排和基因突变的重大风险，可以挑战生物体的福祉并促进发育癌症。有几种细胞机制可以监测基因组的状态，并迅速启动修复机制，以应对DNA损伤，从而将健康的基因组传递给下一代。聚（ADP-核糖）聚合酶-1或PARP-1是对DNA结构中断的主要响应者。 PARP-1具有独特的催化活性，可将ADP-核糖的聚合物作为靶蛋白的翻译后修饰，主要是在PARP-1本身上（自动化）。与DNA断裂结合后，PARP-1活性被“打开”以调节DNA损伤修复途径，从而促进细胞存活。相反，过度的DNA损伤导致PARP-1活性水平升高，导致细胞死亡。因此，PARP-1活性的调节是确定细胞命运的关键因素。此外，PARP-1的抑制剂最近已成为治疗癌症和炎症的有前途的治疗剂。尽管对PARP-1抑制剂的兴趣日益增加，并且发现了在DNA修复，转录调控和凋亡信号传导中发现PARP-1活性的扩展作用，但对PARP-1活性和调节机制的见解很少。该研究计划的长期目标是在分子级建立控制PARP-1活性的机制。 DNA损伤是PARP-1的最有效激活因子。因此，我们选择首先关注PARP-1的DNA依赖性激活的机理。使用X射线晶体学和生化分析的结合，提出的研究将提高我们对PARP-1对DNA损伤的识别以及与染色质质量的PARP-1相互作用的理解（特定的目标1）。这些研究将提供与DNA结合的PARP-1的第一个观点，因此将提供机械见解，这些见解将大大推进PARP研究领域，并对DNA修复和染色质生物学领域产生更大的影响。拟议的工作将证明PARP-1的多个领域如何与夫妇（ADP-ribose）合成与结构特异性DNA结合（特定目标2）合作。 PARP-1活性和调节的详细结构分析将改善PARP-1生物学功能的当前模型，并有可能揭示特定抑制PARP-1活性的新型策略。