Role of PARP-1 in Mediating Inflammatory Gene Transcription

PARP-1 在介导炎症基因转录中的作用

• 批准号: 7324835
• 负责人: MITCHELL C JUNG
• 金额: $ 29.57万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-12-01 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7324835
• 关键词: AIDS Dementia Complex; Acute; Adenosine Diphosphate Ribose; Alzheimer' s Disease; Autoimmune Process; Binding; Cells; Cessation of life; Complex; Data; Development; Disease; Figs - dietary; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Genetic Transcription; Immune response; Inflammation; Inflammatory; Inflammatory Response; Injury; Interleukin-6; Knowledge; Lead; Lung Inflammation; Mediating; Mediator of activation protein; Methods; Molecular; Multiple Sclerosis; Nitric Oxide Synthase; Organ failure; Pathogenesis; Poly Adenosine Diphosphate Ribose; Poly(ADP-ribose) Polymerases; Prevention; Proteins; Recruitment Activity; Regulation; Reporting; Research Personnel; Resistance; Role; Septic Shock; Specificity; Testing; Tissues; design; inhibitor/antagonist; insight; macrophage; mutant; novel; poly(ADP-ribose)polymerase-1, mouse; prevent; programs; promoter; protein protein interaction; response; toll-like receptor 4; transcription factor

Overproduction of inflammatory mediators can lead to major organ failure and damage. The transcription of a defined set of inflammatory genes requires the simultaneous specific DNA-protein and protein-protein interactions in highly ordered complexes called enhanceosome. Poly(ADP-ribose) polymerase-1 (PARP-1) regulates transcription factors responsible for inflammatory gene expression, and importantly PARP-1-deficient (PARP-1-/-) mice were shown to be resistant to inflammatory diseases. The overall HYPOTHESES are that PARP-1 is recruited to the promoter, modifies transcription regulators with poly(ADP-ribose), and mediates recruiting transcription regulators to assemble the enhanceosome for synergistic gene transcription. Our SPECIFIC AIMS are to (1) determine whether PARP- 1 binds the iNOS promoter in a sequence-specific manner; (2) investigate if PARP-1 is recruited to the iNOS promoter by interacting with a sequence-specific transcription factor(s) and increasing its DMA binding activity; (3) investigate whether p42/44MAPK directly or indirectly phosphprylates and induces PARP-1 catalytic activity in response to LPS; (4) determine whether PARP-1 targets transcription regulators with poly(ADP-ribose) for the efficient gene transcription in response to LPS;(5) characterize PARP-1 mediating the recruitment of transcription regulators to the promoters (i) by comparing wild-type and PARP-1-/- cells, (ii) by simultaneously analyzing PARP-1-mediated recruitments, and (iii) of iNOS, TNFalpha, IL-1beta, and IL-6; and (6) definitively determine the specificity of PARP-1 and its catalytic activity on inflammatory gene regulation by expressing wild-type PARP-1 and catalytically inactive PARP-1 mutant in PARP-1-/- cells. The SIGNIFICANCE of the proposed studies is to offer new insights into the mechanism(s) by which PARP-1 may regulate and be regulated to achieve synergistic inflammatory gene transcription. The inflammatory gene transcription is critical for immune responses and for determining the onset of inflammatory diseases such as septic shock, acute lung inflammation, Alzheimer's disease, multiple sclerosis, and HIV-associated dementia.

炎症介质的过度产生可导致主要器官衰竭和损伤。的 一组确定的炎性基因的转录需要同时特异的DNA-蛋白质， 蛋白质-蛋白质相互作用形成高度有序的复合体，称为增强体。聚ADP-核糖 聚合酶-1（PARP-1）调节负责炎性基因表达的转录因子， 重要的是，显示PARP-1缺陷（PARP-1-/-）小鼠对炎性疾病具有抗性。 总体假设是PARP-1被募集到启动子，修饰转录 调节器与聚（ADP-核糖），并介导招募转录调节器组装 用于协同基因转录的增强体。我们的具体目标是（1）确定PARP- 1以序列特异性方式结合iNOS启动子;（2）研究PARP-1是否被募集到iNOS启动子中; 通过与序列特异性转录因子相互作用并增加其DMA结合， 研究p42/44 MAPK是否直接或间接磷酸化并诱导PARP-1的表达 响应LPS的催化活性;（4）确定PARP-1是否靶向转录调节因子， （5）研究了PARP-1介导的LPS诱导的基因转录 （i）通过比较野生型和PARP-1-/-细胞， (ii)通过同时分析PARP-1介导的募集，和（iii）iNOS、TNF α、IL-1 β和 确定PARP-1的特异性及其对炎症基因的催化活性 通过在PARP-1-/-细胞中表达野生型PARP-1和无催化活性的PARP-1突变体来调节。 拟议研究的意义在于为以下机制提供新的见解： PARP-1可以调节和被调节以实现协同炎症基因转录。的 炎症基因转录对于免疫应答和确定炎症的发生是至关重要的。 炎症性疾病如败血性休克、急性肺部炎症、阿尔茨海默病、多发性硬化症、慢性炎症 硬化症和艾滋病相关痴呆症