Identification of longevity targets in C. elegans protein translation mutants

描述(申请人提供)：本奖学金申请中提出的研究解决了衰老和信使核糖核酸翻译领域中一个非常有趣的问题：为什么许多生物，包括线虫线虫，在信使核糖核酸翻译机制的关键组件上存在缺陷，能够长寿并耐受压力？有趣的是，为了应对环境压力，所有真核细胞都关闭了它们的生物合成活动，并启动了一种综合的应激反应。在恢复过程中，某些蛋白质是在一般翻译抑制的背景下产生的。本应用的主要目的是揭示在应激反应中使应激诱导的mRNAs从翻译抑制中逃脱的生化机制，以及参与长寿表型的主动翻译的mRNAs。该项目将检验一种假说，即属于S6激酶(S6K)和/或真核细胞翻译起始因子(EIF)组的关键翻译调控因子可以特异性地介导这种逃逸。重点将放在rsks-1/S6K以及IFE-2/eIF4E上，IFE-2/eIF4E是蠕虫中的五种亚型之一，是一种mRNA5‘-帽结合蛋白。S6K和eIF4E可能都是调节一条途径的核心，该途径介导了应激特异性和长寿翻译。要解决的主要问题是：S6K和eIF4E下游的翻译靶点是什么，需要合成才能在压力和衰老下存活？这个问题将用一系列尖端的核糖体和蛋白质组学方法来解决，使用线虫，线虫作为实验系统。这种模式生物发展迅速，寿命短，而且能够非常容易地通过RNAi敲除基因，因此对衰老研究非常有用。

DESCRIPTION (provided by applicant): The research proposed in this fellowship application addresses an extremely interesting question in the field of aging and mRNA translation: How come many organisms, including the nematode C. elegans defective in key components of the mRNA translational machinery are long-lived and stress resistant? Interestingly, in response to environmental stress, all eukaryotic cells shut down their biosynthetic activity and mount an integrated stress response. During recovery, certain proteins are produced against the backdrop of general translational repression. The main objective of the present application is to uncover the biochemical mechanisms that enable the escape of stress-induced mRNAs from translational repression during the stress response and the actively translated mRNAs involved in the longevity phenotype. The project will examine the hypothesis that key translational regulatory factors belonging to either S6 kinase (S6K) and/or the group of eukaryotic translation initiation factors (eIFs) can specifically mediate this escape. The focus will be on rsks-1/ S6K, as well as on ife-2/eIF4E, one of five isoforms in worms that is an mRNA 5'-cap binding protein. Both S6K and eIF4E may be at the heart of regulating a pathway that mediates stress-specific and longevity translation. The main question to be addressed is: What are S6K and eIF4E downstream translational targets that need to be synthesized in order to allow survival under stress and aging? This question will be addressed with a series of cutting-edge ribonomic and proteomic approaches using the nematode, C. elegans as an experimental system. The model organism is extremely useful for aging research because of its rapid development, short lifespan, and the ability to knock-out genes through RNAi with great ease.