Structural and genetic variability of the proteostasis network in Saccharomyces cerevisiae.

酿酒酵母蛋白质稳态网络的结构和遗传变异。

• 批准号: 440376057
• 负责人: Dr. Matthias Weith
• 金额: --
• 依托单位: Institut für Biochemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2020
• 资助国家: 德国
• 起止时间: 2019-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/440376057?language=en
• 关键词: Structural; genetic; variability; proteostasis; network

Homeostasis of the cellular ensemble of proteins (proteostasis) is a basic requirement for fitness. Sudden or progressive decline of proteostasis can be pathologic but is also commonly observed in aging. We have not identified the exact causes of this deterioration nor do we understand its link to the accumulation of toxic protein species in disease. Proteostasis decline in aging and disease depends on events within the cellular network of mechanisms that affect proteostasis. Moreover, individual aging trajectories likely depend on combined effects of multiple genetic variants on this network.Proteome-wide measurements are rapidly changing the paradigms of how we study complex traits such as proteostasis. Here, we propose to apply modern mass spectrometry-based methods that are able to detect protein functional state changes on a proteome-wide scale to study the robustness of the model organism Saccharomyces cerevisiae to proteotoxic stress caused by heat. Application of a recently developed structural proteomics method based on limited proteolysis (“LiP-MS”) will allow us to study proteostasis mechanisms and heat stress robustness as a network by simultaneous monitoring of multiple cellular processes. We aim to detemine the genetic foundation of robustness to heat stress based on a panel of genetically different yeast strains. Phenotypic measurements will be combined with information on the individual genetic, transcriptional and proteomic make-up. These data will allow us to relate individual genetic determinants of heat stress robustness to the functional state of regulatory modules. By means of induction, we will chart the network of mechanisms that underlies heat stress robustness, whereas mechanistic hypotheses will be deduced from network information and molecular data.In addition to the investigation of inter-individual differences in thermotolerance traits, we will also determine the influence of prior stress experience. Yeast can acquire thermotolerance following mild stress pre-treatment. We will assess the overlap of changes due to experience with inter-individual determinants of heat stress robustness. Furthermore, we will study the genetic requirements for the establishment of heat stress memory.In sum, this study will take a network-oriented approach combined with encompassing proteomics methods to elucidate basic processes underlying failure and defense of cellular proteostasis. In-depth investigation of mechanistic hypotheses deduced from proteome-wide measurements will be carried out using detailed molecular biology experiments. Thereby, the ability of our approach to identify inter-connected determinants of stress robustness will be demonstrated.

蛋白质细胞群的稳态（蛋白质稳态）是健康的基本要求。蛋白质稳态的突然或进行性下降可能是病理性的，但也常见于衰老过程中。我们尚未确定这种恶化的确切原因，也不了解其与疾病中有毒蛋白质种类积累的联系。衰老和疾病导致的蛋白质稳态下降取决于影响蛋白质稳态的细胞机制网络内的事件。此外，个体衰老轨迹可能取决于多个遗传变异对该网络的综合影响。蛋白质组范围的测量正在迅速改变我们研究蛋白质稳态等复杂性状的范式。在这里，我们建议应用基于现代质谱的方法，能够在蛋白质组范围内检测蛋白质功能状态的变化，以研究模型生物酿酒酵母对热​​引起的蛋白质毒性应激的稳健性。应用最近开发的基于有限蛋白水解（“LiP-MS”）的结构蛋白质组学方法将使我们能够通过同时监测多个细胞过程来研究蛋白质稳态机制和热应激稳健性作为网络。我们的目标是根据一组基因不同的酵母菌株来确定对热应激的鲁棒性的遗传基础。表型测量将与个体遗传、转录和蛋白质组组成的信息相结合。这些数据将使我们能够将热应激稳健性的个体遗传决定因素与调节模块的功能状态联系起来。通过归纳法，我们将绘制出热应激鲁棒性的机制网络，并从网络信息和分子数据中推导出机制假设。除了研究耐热性特征的个体间差异外，我们还将确定先前应激经历的影响。经过轻微应激预处理后，酵母可以获得耐热性。我们将根据热应激稳健性的个体间决定因素的经验来评估变化的重叠。此外，我们将研究建立热应激记忆的遗传要求。总之，本研究将采用面向网络的方法结合蛋白质组学方法来阐明细胞蛋白质稳态失败和防御的基本过程。将使用详细的分子生物学实验对从全蛋白质组测量中推导出来的机制假设进行深入研究。因此，我们的方法能够识别压力鲁棒性的相互关联的决定因素。