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

项目摘要/摘要 这项拨款提案寻求确定第一个已知的细胞骨架特异性应激反应和 这一途径的潜在调节者。之前在酵母菌上的研究发现，与年龄相关的 细胞骨架功能及其对寿命的影响，而最近在线虫中的工作发现HSF-1在 在细胞骨架完整性中起到保护作用。结合这些发现，我们建议将这两个特征 在多细胞模式生物中，全身性和组织特异性细胞骨架随年龄的变化而下降，C. 优雅女装。接下来，我们想要确定与HSF-1协同或独立于HSF-1在 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 应激反应本身会随着年龄的增长而下降，就像许多应激反应途径一样--包括线粒体， 内质网和胞浆--以及异位激活是否能挽救细胞骨架功能 年事已高。 我们将使用生化和成像方法来验证我们的假设。我们建议研究 通过使用LifeAct对肌动蛋白进行成像来实现细胞骨架功能。此外，我们将提纯组织中的肌动蛋白- 并研究它们在成虫不同阶段的功能。最后，我们建议 进行400个转录因子的候选筛选，以确定细胞骨架功能的新调节因子。 在这里，内吞作用和肌肉纤维的组织将被用作细胞骨架完整性的读数和 功能。在这项研究的高潮，我们将把细胞骨架的应激反应描述为 年龄的作用。展望未来，这将开辟令人兴奋的研究途径，继续制定 细胞骨架应激反应的机制途径，以及确定这一保守性 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.