Defining the programmed proteome rejuvenation underlying gametogenesis

定义配子发生背后的程序化蛋白质组复兴

• 批准号: 10471317
• 负责人: Gloria Ann Brar
• 金额: $ 57.45万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10471317
• 关键词: Age; Aging; Animal Model; Atlases; Biological Assay; Cell Aging; Cell physiology; Cells; Cellular Structures; Collaborations; Complex; Coupled; Data; Disease; Ensure; Excision; Exclusion; Exhibits; Functional disorder; Future Generations; Gametogenesis; Germ Cells; Goals; Housekeeping; Incidence; Inherited; Link; Longevity; Mass Spectrum Analysis; Measurement; Measures; Meiosis; Mitotic; Molecular; Organism; Outcome Study; Pathway interactions; Physiologic pulse; Physiology; Population; Process; Production; Proteins; Proteome; Proteomics; Quality Control; Regulation; Rejuvenation; Ribosomes; Saccharomyces cerevisiae; Saccharomycetales; Sexual Reproduction; Specificity; Sum; Surveys; Testing; Time; Yeasts; Youth; age related; aged; base; cell age; cell injury; cell type; combat; cost; experience; fitness; insight; interest; molecular marker; multicatalytic endopeptidase complex; oxidation; portability; precursor cell; prevent; programs; protein aggregation; protein complex; protein degradation; protein function; proteostasis; screening; superoxide dismutase 1; transcription factor; transcriptome

ABSTRACT The integrity of a cell depends on the quality of its components. These components include proteins, which are responsible for executing most cellular functions, many through organization into stable complexes. A hallmark of aged cells is the breakdown of protein integrity, or proteostasis, which results from the damage to key proteins and complexes over time, leading to more accumulated damage, and ultimately cell dysfunction. The specific proteome components that are most susceptible to damage and that drive its accumulation remain unclear, but the survival of future generations depends on protection of one cell type—gametes—from inheriting damaged components from their precursor cell. During gametogenesis in the simple budding yeast, as a precursor cell is differentiated into gametes, we observe the degradation of many cellular structures and proteins, followed by their resynthesis and reorganization. This cellular restructuring is associated with an active rejuvenation program that allows equivalently young gametes to be produced from old or young precursor cells. The mechanisms that contribute to this natural rejuvenation program are not known, but it can be recapitulated by exogenous expression of a meiotic transcription factor in aged mitotic cells, suggesting that it is portable. Gametogenesis in yeast thus offers the opportunity to watch as the cell shows us what proteins and complexes it needs to reset and reorganize to ensure cellular youth, and the mechanisms it uses to achieve this. Of particular interest are proteins of basal or “housekeeping” function, including the ribosome, which are long- lived in mitotic cells, but degraded and replaced at great energetic cost during gametogenesis. We also observe reorganization of abundant housekeeping complexes, including the proteasome, during gametogenesis, and aggregation of some proteins, including superoxide dismutase 1 (Sod1), at the time of their degradation. Together, these observations suggest that yeast cells remodel their proteome during gametogenesis as a quality control measure. Here, we propose to identify the key set of cellular components, with a focus on “housekeeping” proteins, that are reset as gametes are created from precursor cells. We use a proteomic approach to globally define changes to protein complexes during gamete construction, and specifically determine proteasome remodeling and activity over time. We investigate the links between aggregation of proteins, including Sod1, and protein oxidation and degradation. Finally, we identify the specific degradation mechanisms that drive key aspects of proteome remodeling and test their necessity for gamete rejuvenation and ability to drive lifespan extension when exogenously activated. This project will build an atlas to reveal the proteins and complexes that are important enough for young cell identity to warrant the energetic cost of resetting them during gamete formation, as well as those that may be toxic enough to warrant their active removal. By identifying and manipulating the specific pathways used by gametes to selectively remodel their proteome, we will find strategies that can be co-opted to combat and prevent age-associated cellular damage.

摘要 电池的完整性取决于其组件的质量。这些成分包括蛋白质， 负责执行大多数细胞功能，许多通过组织成稳定的复合体。一个标志 衰老细胞的主要特征是蛋白质完整性的破坏，或蛋白质稳态，这是由关键蛋白质的损伤引起的 随着时间的推移和复合物，导致更多的累积损伤，最终细胞功能障碍。具体 最容易受到损伤并促使其积累的蛋白质组成分尚不清楚， 后代的生存取决于保护一种细胞类型-配子-免受遗传损伤。 从它们的前体细胞中分离出来。在简单芽殖酵母的配子发生过程中，作为前体细胞， 在分化为配子的过程中，我们观察到许多细胞结构和蛋白质的降解， 它们的重新合成和重组。这种细胞重组与一个积极的复兴计划有关 这使得同样年轻的配子可以从年老或年轻的前体细胞中产生。的机制 有助于这种自然复兴计划是未知的，但它可以概括为外源 在衰老的有丝分裂细胞中减数分裂转录因子的表达，表明它是可携带的。 因此，酵母中的配子发生提供了观察细胞向我们展示什么蛋白质和 它需要重置和重组以确保细胞年轻的复合体，以及它用来实现这一目标的机制。 特别令人感兴趣的是具有基础或“管家”功能的蛋白质，包括核糖体，它们是长链的。 生活在有丝分裂细胞中，但在配子发生过程中以巨大的能量代价降解和替换。我们还观察 在配子发生过程中重组丰富的管家复合物，包括蛋白酶体，以及 某些蛋白质，包括超氧化物歧化酶1（SOD 1）在降解时的聚集。 总之，这些观察结果表明，酵母细胞在配子发生过程中重塑其蛋白质组作为一种质量 控制措施在这里，我们建议确定关键的一组细胞成分，重点是“管家” 这些蛋白质在配子由前体细胞产生时被重置。我们使用蛋白质组学方法， 定义配子构建过程中蛋白质复合物的变化，并特别确定蛋白酶体 随着时间的推移重塑和活动。我们研究了蛋白质聚集（包括Sod 1）和 蛋白质氧化降解。最后，我们确定了驱动关键的特定退化机制， 蛋白质组重塑的各个方面，并测试它们对配子恢复活力和延长寿命的能力的必要性 外生激活时的延伸。该项目将建立一个图谱，以揭示蛋白质和复合物， 对于年轻细胞的身份来说是足够重要的，以保证在配子期间重置它们的能量成本 形成，以及那些可能是有毒的，足以保证他们的积极清除。通过识别和 操纵配子使用的特定途径来选择性地重塑它们的蛋白质组，我们将找到策略， 可以用来对抗和预防与年龄相关的细胞损伤。