Oxidative Stress, Mitochondrial Dysfunction and Aging

项目描述（由申请人提供）：本项目包括三个研究项目和一个行政核心。我们的工作假设提出(a)衰老组织表现出线粒体生成ROS的增加，导致(b) ROS产生的锁定循环，放大氧化应激；(c)稳定由ROS引起的应激调节因子的稳态变化(d)导致慢性应激状态和组织功能的进行性下降。我们的PP关注的是这些ros介导的变化的后果。P1 （JP）研究了p38 MAPK及其靶向转录因子和线粒体因子在衰老肝脏中的功能，以及对3-NPA、鱼烯酮和抗霉素A、线粒体复合物I、II和III抑制剂产生ROS的信号的响应。P1。作为分包商，WRW将确定3- npa产生ROS的地点，并与所有3个项目进行机械互动。P2 （SM）研究衰老如何影响APE1在核和mtDNA修复中的作用；负Ca++反应元件（nCaRE）在Ca++反应基因表达调控中的作用，改变转基因小鼠细胞核和APE水平如何影响细胞对ROS的反应；应激诱导的APE1修饰的年龄依赖性调节是否影响衰老细胞的剪切和氧化应激反应。P3 （IB）研究了8氧鸟嘌呤DNA糖基化酶（OGG1）对mtDNA修复的调节，以及年龄相关的线粒体功能下降是否由OGGI的表达/活性、翻译后修饰和/或区隔化变化引起的DNA修复受损/低效引起。这三个项目都有助于我们共同的PP目标，了解线粒体功能障碍产生的ROS的影响，以及蛋白质修饰如何影响应激反应的变化和每个项目中研究的基因活性。行政核心A将协助PI完成行政任务并协调动物使用。程序性互动包括：积极合作，共享组织以减少动物成本；积极的系列讨论会；并且经常就实验设计、方法和数据分析进行咨询。通过在相同和不同的实验系统中引入一系列不同的方法来承担各种年龄相关的应激反应变化，我们将更清楚地发现影响正常组织功能随年龄下降的共同调控主题。

DESCRIPTION (provided by applicant): This Program Project consists of three research projects and an Administrative Core. Our working hypothesis proposes that (a) aging tissues exhibit an increase in mitochondrially generated ROS causing (b) a locked-in cycle of ROS production that amplifies oxidative stress and (c) stabilizes the homeostatic changes in regulators of the stress caused by the ROS (d) that result in a state of chronic stress and progressive decline in tissue function. Our PP is focused on the consequences of these ROS-mediated changes. P1 (JP) investigates the function of p38 MAPK, its targeted transcription factors and mitochondriai factors in the aged liver and in response to signals due to ROS generation by 3-NPA, rotenone, and antimycin A, inhibitors of mitochondrial complexes I, II and III. P1. A subcontractor, WRW will identify the site of ROS generated by 3-NPA and interact mechanistically with all 3 projects. P2 (SM) investigates how aging affects the role of APE1 in nuclear and mtDNA repair; the role of the negative Ca++ response element (nCaRE) in regulation of expression of Ca++ - responsive genes, how changing the nuclear and APE levels in transgenic mice affects the cellular response to ROS; whether the age-dependent modulation of stressinduced APE1 modification affects the shear and oxidative stress responses in aged cells. P3 (IB) investigates the regulation of mtDNA repair by 8 oxo-guanine DNA glycosylase (OGG1), and whether the age-associated decline in mitochondrial function is caused by impaired/inefficient DNA repair due to changes in expression/activity, posttranslational modification and/or compartmentalization of OGGI. All three projects contribute to our shared PP aims, to understand the effects of ROS generated by mitochondrial dysfunction and how protein modifications may affect changes in stress responses and the activity of their genes investigated in each project. The Administrative Core A will assist the PI in administrative tasks and coordinate animal use. Programmatic interactions include: active collaborations, sharing tissues to reduce animal costs; an active seminar series; and frequent consultations as to experimental design, methodology, and data analysis. By bringing a range of different approaches to bear on various age-related changes in stress response in the same and different experimental systems, we will more clearly discern common regulatory themes that affect the decline in proper tissue functions with age.