The Cytoskeletal Stress Response: A Novel Facet to Protecting Cell Integrity during Aging

项目概要/摘要 这项拨款提案旨在确定第一个已知的细胞因子特异性应激反应， 这条通路的潜在调节剂。以前在酵母中的工作已经确定了与年龄相关的 细胞骨架功能及其在寿命中的意义，而最近的工作在C。秀丽线虫已经鉴定出HSF-1， 介导细胞骨架完整性的保护作用。结合这些发现，我们建议将两者 系统性和组织特异性细胞骨架下降作为年龄的函数在多细胞模型生物，C。 优雅接下来，我们想确定在HSF-1中协同或独立工作的关键参与者， 在压力和老化下保护细胞骨架的完整性。最后，我们可以确定细胞骨架是否 压力反应本身随着年龄的增长而下降，就像许多压力反应途径一样-包括线粒体， 内质网和细胞质-以及异位激活是否可以挽救细胞骨架功能， 高龄 我们将采用生物化学和成像方法来验证我们的假设。我们建议研究 细胞骨架功能的肌动蛋白成像使用LifeAct。另外，我们将纯化组织中的肌动蛋白- 具体的方式，并研究其功能的蠕虫在不同阶段的成年。最后，我们建议 对400个转录因子进行候选筛选，以鉴定细胞骨架功能的新调节因子。 在这里，肌纤维的内吞作用和组织将用作细胞骨架完整性的读数， 功能在这项研究的高潮，我们将有特点的细胞骨架应激反应作为一个 年龄的函数。展望未来，这将开辟令人兴奋的研究途径，继续绘制 细胞骨架应激反应的机制途径，以及确定这种保守性 在哺乳动物和疾病模型中的机制。

Project Summary/Abstract This grant proposal seeks to identify the first known cytoskeleton-specific stress response and the potential regulators of this pathway. Previous work in yeast has identified an age-associated decline in cytoskeletal function and its implications in lifespan, while recent work in C. elegans have identified HSF-1 in mediating a protective role in cytoskeletal integrity. Marrying these findings, we propose to characterize both systemic and tissue-specific cytoskeletal decline as a function of age in the multicellular model organism, C. elegans. Next, we would like to identify the key players working both in synergy or independently of HSF-1 in protecting cytoskeletal integrity under stress and aging. Finally, we can determine whether the cytoskeletal stress response itself declines over age, as many stress response pathways do – including mitochondrial, endoplasmic reticulum, and cytoplasmic – and whether ectopic activation can rescue cytoskeletal function in advanced age. We will employ both biochemical and imaging methods to test our hypotheses. We propose to study cytoskeletal function by imaging of actin using LifeAct. In addition, we will purify actin proteins in a tissue- specific manner and study their function in worms at various stages of adulthood. Finally, we propose to perform a candidate screen of 400 transcription factors to identify novel regulators of cytoskeletal function. Here, endocytosis and organization of muscle fibers will be used as a readout for cytoskeletal integrity and function. At the culmination of this study, we will have characterized the cytoskeletal stress response as a function of age. Moving forward, this will open exciting avenues of research in continuing to map out the mechanistic pathway of the cytoskeletal stress response, as well as identifying the conservation of this mechanism in mammals and disease models.