Mechanistic dissection of eukaryotic protein biogenesis and degradation pathways

真核蛋白质生物发生和降解途径的机制剖析

• 批准号: 10623737
• 负责人: Vladimir Denic
• 金额: $ 57.59万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10623737
• 关键词: 2019-nCoV; 3-Dimensional; Age; Amino Acids; Area; Autophagocytosis; Autophagosome; Biochemical; Biogenesis; Cell Culture Techniques; Cells; Cues; Degradation Pathway; Dissection; Encapsulated; Endoplasmic Reticulum; Genetic Screening; Genomics; Heat-Shock Response; Human; Intracellular Membranes; Link; Microscopy; Molecular Chaperones; Nerve Degeneration; Organelles; Peptide Elongation Factor 1; Pharmaceutical Preparations; Phenotype; Physiological; Process; Production; Proteins; Quality Control; Shapes; Signal Transduction; Starvation; Stress; System; Testing; Theoretical model; Ubiquitin; Yeasts; biological adaptation to stress; interest; multicatalytic endopeptidase complex; mutant; peroxisome; preclinical efficacy; protein degradation; protein structure function; proteostasis; receptor; reconstitution

Abstract My lab currently has two main areas of interest: 1) a new chaperone system (eFOLD) that enables biogenesis of eukaryotic translation elongation factor 1 alpha (eEF1A); 2) endoplasmic reticulum (ER) and peroxisome degradation by selective autophagy. Errors in eEF1A biogenesis result in rapid degradation by the ubiquitin-proteasome system (UPS) thus making protein degradation a natural link between the two areas. Both eFOLD and selective autophagy are controlled by distinct stress responses (e.g. heat shock vs. amino acid starvation) but jointly serve as effectors of protein homeostasis (proteostasis). Both areas raise similar questions regarding substrate selectivity: How does a specific eFOLD chaperone co-translationally recognize an aggregation-prone region of eEF1A nascent chains? How is terminally misfolded eEF1A recognized for degradation by the UPS? What signals on damaged or unwanted organelles are detected by specific autophagy receptors to orchestrate encapsulation of organelle targets into autophagosomes? To answer these questions, we are dissecting biogenesis and degradation mechanisms that select substrates of grossly different sizes, respond to distinct physiological cues, and have widely different temporal dynamics. Using yeast and human cell culture in parallel, we are exploring conserved aspects of eFOLD and selective autophagy mechanisms shared by each species, as well as species-specific adaptations. Broadly speaking, our projects spawn from identification of missing factors by genetic screening or biochemical purification but all seek a deep mechanistic understanding of mutant phenotypes through biochemical reconstitution with purified components and protein structure-function analysis. Along this path, we iteratively test our hypotheses by genomics, quantitative cell microscopy, and theoretical modeling approaches.

抽象的 我的实验室目前有两个主要兴趣领域：1）一种新的伴侣系统（eFOLD），可以实现生物发生 真核翻译延伸因子 1 α (eEF1A) 内质网 (ER) 和过氧化物酶体； eEF1A 生物合成中的错误导致选择性自噬降解。 泛素-蛋白酶体系统（UPS）从而使蛋白质降解成为这两个领域之间的自然联系。 eFOLD 和选择性自噬均由不同的应激反应控制（例如热休克与氨基 酸饥饿），但共同充当蛋白质稳态（蛋白质稳态）的影响者。 关于底物选择性的问题：特定的 eFOLD 伴侣如何共翻译识别 eEF1A 新生链的易聚集区域如何被识别为末端错误折叠的 eEF1A？ 特定的 UPS 检测到受损或不需要的细胞器上的哪些信号？ 自噬受体协调将细胞器靶标封装到自噬体中？ 这些问题，我们正在剖析生物发生和降解机制，这些机制选择了粗略的底物 不同的尺寸，对不同的生理信号做出反应，并且具有截然不同的时间动态。 酵母和人类细胞培养并行，我们正在探索 eFOLD 的保守方面和选择性 每个物种共有的自噬机制，以及物种特异性的适应。 我们的项目源于通过基因筛选或生化纯化来识别缺失的因子，但是 所有人都寻求通过生化重建对突变表型进行深入的机制理解 沿着这条路径，我们迭代地测试我们的纯化成分和蛋白质结构功能分析。 通过基因组学、定量细胞显微镜和理论建模方法提出假设。

项目成果

Autophagy prevents runaway meiotic divisions.

自噬可防止减数分裂失控。

• 发表时间: 2020
• 影响因子: 13.3
• 作者: Wang, Fei;Denic, Vladimir;Lacefield, Soni
• 通讯作者: Lacefield, Soni

A TRCky TA protein delivery service snubs the UPS.

TRCky TA 蛋白质递送服务冷落了 UPS。

• 发表时间: 2021-05-03
• 作者: McQuown, Alexander J;Reif, Dvir;Denic, Vladimir
• 通讯作者: Denic, Vladimir

Defining the Essential Function of Yeast Hsf1 Reveals a Compact Transcriptional Program for Maintaining Eukaryotic Proteostasis.

定义酵母 Hsf1 的基本功能揭示了维持真核蛋白质稳态的紧凑转录程序。

• DOI: 10.1016/j.molcel.2016.05.014
• 发表时间: 2016-07-07
• 期刊: Molecular cell
• 影响因子: 16
• 作者: Solís EJ;Pandey JP;Zheng X;Jin DX;Gupta PB;Airoldi EM;Pincus D;Denic V
• 通讯作者: Denic V

Putting Your Best Egg Forward.

全力以赴。

• 发表时间: 2018
• 影响因子: 11.8
• 作者: Patil, Christopher K;Denic, Vladimir
• 通讯作者: Denic, Vladimir

A Zpr1 co-chaperone mediates folding of eukaryotic translation elongation factor 1A via a GTPase cycle.

Zpr1 共伴侣通过 GTP 酶循环介导真核翻译延伸因子 1A 的折叠。

• 发表时间: 2023-09-07
• 影响因子: 16
• 作者: McQuown, Alexander J;Nelliat, Anjali R;Reif, Dvir;Sabbarini, Ibrahim M;Membreno, Britnie Santiago;Wu, Colin Chih;Denic, Vladimir
• 通讯作者: Denic, Vladimir