Multidimensional mapping of proteome changes and mechanisms underlying yeast replicative aging

项目摘要 衰老研究的主要目标之一是了解细胞如何随着时间的推移逐渐退化。几十年的 分子和细胞生物学的工作已经发现了细胞衰老的几个保守特征，包括 线粒体功能障碍和空泡/溶酶体缺陷。这些老化的标志通常是一个一个地研究的。 时间和离开的原因和它们之间的联系在很大程度上是未知的。触发因素和联系是什么 在衰老的不同标志中逐步达到顶峰的分子事件之间的关系？没有大局观 关于蛋白质组中每种蛋白质的变化，我们所能做的最好的事情往往是猜测和尝试， 这些特征之间的联系为了填补空白，我们制定并实施了一项新的 高通量成像方法，系统地跟踪每一个蛋白质的命运，包括其表达， 在萌芽的复制老化过程中，这些变化的锡永、定位、聚集和时间（4D命运图） 酵母正在进行的4D命运作图工作已经证明了其在揭示新的年龄相关分子中的价值。 这是众所周知的衰老标志的主要原因。一个典型的例子， 一个初步的结果是，与年龄相关的Tom 70减少是线粒体衰老的关键事件。 过度表达Tom 70可以防止与年龄相关的线粒体生物合成缺陷和空泡酸化。 fification--细胞老化的两个标志。这项提案的目的是深入研究分子机制- 这些新的Tom 70功能的分析，这将作为一个例子，使用蛋白质组命运映射，以揭示 未知原因和老化标志的联系（目标1和目标2）。我们的初步研究表明 我们将在本资助周期内完成整个酵母蛋白质组图谱的绘制。完成 命运绘图（目标3）不仅将通过揭示额外的机制来补充其他两个目标， 有助于线粒体和液泡的老化（目标1和目标2），但也填补了所有其他 老化的标志我们的初步结果证明了这一点，即正在进行的命运绘图提供了一个不确定的， 老化过程中Tom 70减少的预期机制。我们希望揭示Tom 70的其他机制 减少和线粒体/空泡老化时，命运映射涵盖更多的蛋白质。这个项目将 推进我们对衰老的理解的深度和广度，并提供命运地图的例子。 (Aim 3）可用于全面了解衰老标志的多因素原因（例如，线粒体 Aim 1和Aim 2中的气泡和空泡缺陷）。研究界挖掘蛋白质组衰老的4D图谱 将系统地揭示以前未知的机制和不同衰老标志之间的联系。 该项目的完成将有利于研究人员寻求不同的触发因素和联系， 老化的标志

PROJECT SUMMARY One of the major goals of aging research is to understand how cells gradually degenerate overtime. Decades of work by molecular and cell biologicals have discovered several conserved hallmarks of cellular aging, including mitochondrial dysfunction and vacuole/lysosome defects. These aging hallmarks are usually studied one at a time and left the causes and connections between them largely unknown. What are the triggers and connections between molecular events that step-by-step culminate at the different hallmarks of aging? Without the big picture of how each protein changes in a proteome, often the best we can do is guess and try when it comes to the causes and connections between these hallmarks. To fill the gaps, we have developed and implemented a new high-throughput imaging method to systematically track the fate of each individual protein, including its expres- sion, localization, aggregation, and timing of these changes (4D fate map), during the replicative aging of budding yeast. The ongoing 4D fate mapping effort has demonstrated its value in revealing novel age-related molecular signatures and the primary causes for the well-known hallmarks of aging. A representative example from our pilot fate mapping effort is that the age-associated reduction of Tom70 is a key event of mitochondrial aging. Overexpressing Tom70 can prevent the age-associated defects in mitochondrial biogenesis and vacuole acidi- fication--two hallmarks of cellular aging. The goal of this proposal is to take a deep dive into the molecular mech- anisms of these novel Tom70 functions, which will serve as an example of using proteome fate mapping to reveal unknown causes and connections of aging hallmarks (Aim 1 and Aim 2). As our preliminary study has cleared the technical barriers, we will finish the mapping of entire yeast proteome within this grant cycle. The completion of fate mapping (Aim 3) will not only complement the other two aims by unfolding additional mechanisms that contribute to the aging of mitochondria and vacuole (Aim 1 and Aim 2) but also fill the gaps between all other aging hallmarks. This is exemplified by our preliminary results that the ongoing fate mapping provided an unex- pected mechanism of Tom70 reduction during aging. We expect to reveal additional mechanisms of Tom70 reduction and mitochondrial/vacuolar aging when fate mapping covers more proteins. Together, this project will advance both the depth and breadth of our understanding of aging and provide examples of how the fate map (Aim 3) can be used to comprehensively understand the multifactorial causes of aging hallmarks (e.g., mitochon- drial and vacuole defects in Aim 1 and Aim 2). Mining the 4D map of proteome aging by the research community will systematically unveil previous unknown mechanisms and connections between different aging hallmarks. The completion of this project will benefit researchers seeking the triggers and connections between different aging hallmarks.