CAREER: Integrating Chemical Biology Methods and RNA Virus Models to Elucidate How the Metazoan Proteostasis Network Modulates Protein Evolutionary Landscapes

职业：整合化学生物学方法和 RNA 病毒模型，阐明后生动物蛋白质稳态网络如何调节蛋白质进化景观

• 批准号: 1652390
• 负责人: Matthew Shoulders
• 金额: $ 103.46万
• 依托单位: Massachusetts Institute of Technology
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-15 至 2022-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1652390&HistoricalAwards=false
• 关键词: CAREER; Integrating; Chemical; Biology; Methods

What are the mutations, harmful or beneficial, that may occur in a protein? The answers to this fundamental question underpin the ability to understand and predict evolution. While trying to study this question by introducing mutations in the lab, it was learned that mutations very often result in the production of protein variants that do not fold correctly in space. In the living cell, there are several systems that assist the protein folding process (known as the cellular proteostasis network), and they help some of the mutant proteins fold correctly. By perturbing the proteostasis systems, the investigator hopes to learn more about the mutations that actually happen during protein evolution. This project will use the influenza viruses, which can evolve rapidly in the lab and chemical probes that can turn on and off various parts of the proteostasis machinery. The research is linked to developing and testing an outreach program targeted at the local homeschooled student community. Opportunities offered include high school student internships related to the research, chemistry demonstration shows, and the development and testing of biomolecule models for take-home experiments that teach students about evolution and protein folding.This research is uncovering how metazoan proteostasis mechanisms beyond the HSP90 chaperone influence protein evolution at the molecular level. A battery of custom-designed chemical biology tools is used to perturb the composition and activities of key components of the metazoan proteostasis network with high precision using small molecules. The effects of these different proteostasis environments on protein evolutionary trajectories are studied by leveraging rapidly evolving RNA viruses propagating in metazoan cells, with a focus on influenza A. The highly interdisciplinary experimental approach encompasses deep mutational scanning of individual viral proteins, and optimized sequencing, modeling, and biophysical analysis strategies to enable quantitative assessment of the results. The outcomes will be a new appreciation for the complex interplay between the proteostasis network and client protein evolution, with important implications for understanding and predicting viral evolution at the molecular level. Testing roles of the proteostasis network in buffering protein evolution will also elucidate previously unknown functions of specific proteostasis network components. Thus, fundamental science contributions will impact fields ranging from virology and evolutionary biology to biophysics.

蛋白质中可能发生的有害或有益的突变是什么？这个基本问题的答案是理解和预测进化的能力的基础。在试图通过在实验室中引入突变来研究这个问题的同时，据了解，突变通常会导致产生无法正确折叠在空间中的蛋白质变体。在活细胞中，有几个系统可以帮助蛋白质折叠过程（称为细胞蛋白抑制网络），它们有助于某些突变蛋白正确折叠。通过扰动蛋白质的系统，研究人员希望更多地了解蛋白质进化过程中实际发生的突变。该项目将使用流感病毒，该病毒可以在实验室中迅速发展，化学探针可以打开和关闭蛋白质机械的各个部分。这项研究与制定和测试针对当地家庭教育学生社区的外展计划有关。提供的机会包括与研究相关的高中生实习，化学示范表明以及用于向学生实验的生物分子模型的开发和测试，这些实验向学生传授了进化和蛋白质折叠的知识。这项研究正在发现超出HSP90蛋白结构蛋白的蛋白质抑制剂机制如何影响HSP90分子蛋白在分子水平上影响蛋白质的蛋白质进化。一系列定制设计的化学生物学工具用于扰动使用小分子高精度的后生动物蛋白质量网络的关键组成部分的组成和活动。通过利用快速进化的RNA病毒在后生动物细胞中传播的RNA病毒，研究了这些不同的蛋白质量环境对蛋白质进化轨迹的影响，重点介绍了流感A。高度跨学科的实验方法涵盖了对单个病毒蛋白的深度突变扫描的深度突变扫描。结果将是对蛋白抑制网络和客户蛋白演化之间复杂相互作用的新欣赏，这对理解和预测分子水平的病毒演化具有重要意义。蛋白质抗体网络在缓冲蛋白演化中的测试作用还将阐明特定蛋白质的网络组件的先前未知功能。因此，基本科学贡献将影响从病毒学和进化生物学到生物物理学等领域。

项目成果

Host proteostasis modulates influenza evolution

• DOI: 10.7554/elife.28652
• 发表时间: 2017-09
• 期刊: eLife
• 影响因子: 7.7
• 作者: Angela M. Phillips;Luna O Gonzalez;Emmanuel E. Nekongo;A. I. Ponomarenko;Seán McHugh;Vincent L. Butty;S. Levine;Yu-Shan Lin;L. Mirny;M. Shoulders

HSF1 Activation Can Restrict HIV Replication

• DOI: 10.1021/acsinfecdis.0c00166
• 发表时间: 2020-07-10
• 期刊: ACS INFECTIOUS DISEASES
• 影响因子: 5.3
• 作者: Nekongo, Emmanuel E.;Ponomarenko, Anna, I;Shoulders, Matthew D.
• 通讯作者: Shoulders, Matthew D.