Imaging large macromolecular complexes inside cells with electron cryotomography

使用电子冷冻断层扫描对细胞内的大分子复合物进行成像

• 批准号: 10013429
• 负责人: GRANT J JENSEN
• 金额: $ 8.17万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10013429
• 关键词: 3-Dimensional; Address; Architecture; Automation; Automobile Driving; Award; Biological Models; Biological Process; Categories; Cell division; Cells; Cellular Structures; Cellular biology; Cryoelectron Microscopy; Cytoskeleton; DNA; Development; Disease; Electrons; Filament; Funding; Future; Goals; Grain; HIV; Image; Infection; Ions; Laboratories; Life; Location; Macromolecular Complexes; Measurement; Molecular; Public Health; Research Project Summaries; Structure; System; Testing; Work; cryogenics; falls; in vivo; method development; microbial; pathogenic bacteria; structural biology; technology development; therapeutic development

Project Summary Research in my laboratory seeks to address the question of what large macromolecular complexes exist inside cells and how their function is determined by their structure and location. To accomplish this goal, we use electron cryotomography (ECT) to image native cellular structures in 3D inside intact cells, carrying out structural biology in vivo. Projects in the lab fall into three categories: microbial cell biology, structural biology of HIV, and cryo-EM methods development. The HIV work is complementary to our other projects in that it drives and is driven by the same technology development, but it is funded separately. In microbial cell biology, in the next five years we aim to advance our understanding of: (1) the architectures and relationships of bacterial secretion systems; (2) the mechanisms of cell division across all three kingdoms of life; and (3) the bacterial cytoskeleton and DNA-segregating filaments. Driving these projects, in cryo-EM methods development, we aim to: (1) expand the throughput and automation of ECT; (2) apply cryogenic focused ion beam-milling to expand the reach of ECT to any cell, regardless of size; and (3) use our ECT measurements to inform the development of realistic coarse-grained models of biological processes to develop and test mechanistic hypotheses.

项目概要 我实验室的研究旨在解决内部存在哪些大分子复合物的问题 细胞以及它们的功能如何由它们的结构和位置决定。为了实现这个目标，我们使用 电子冷冻断层扫描 (ECT) 对完整细胞内的天然细胞结构进行 3D 成像， 体内结构生物学。实验室的项目分为三类：微生物细胞生物学、结构生物学 HIV 和冷冻电镜方法开发。艾滋病毒工作与我们的其他项目是互补的，因为它 驱动和受同一技术开发驱动，但资金是分开的。在微生物细胞生物学中， 在未来五年中，我们的目标是加深对以下方面的理解：（1） 细菌分泌系统； (2) 生命三个王国的细胞分裂机制；和（3） 细菌细胞骨架和 DNA 分离丝。通过冷冻电镜方法推动这些项目 开发时，我们的目标是：（1）扩大ECT的吞吐量和自动化程度； (2)应用低温聚焦离子 光束铣削将 ECT 的覆盖范围扩大到任何细胞，无论细胞大小； (3) 使用我们的 ECT 测量 为开发和测试生物过程的现实粗粒度模型提供信息 机制假设。

项目成果

In Situ Imaging and Structure Determination of Biomolecular Complexes Using Electron Cryo-Tomography.

使用电子冷冻断层扫描对生物分子复合物进行原位成像和结构测定。

• DOI: 10.1007/978-1-0716-0966-8_4
• 发表时间: 2020-12-27
• 期刊: Methods in molecular biology
• 作者: M. Kaplan;William J. Nicolas;Wei Zhao;S. Carter;L. Metskas;G. Chreifi;D. Ghosal;G. Jensen
• 通讯作者: G. Jensen

FtsEX-mediated regulation of the final stages of cell division reveals morphogenetic plasticity in Caulobacter crescentus.

FtsEX 介导的细胞分裂最后阶段的调节揭示了新月柄杆菌的形态发生可塑性。

• 发表时间: 2017-09
• 影响因子: 4.5
• 作者: Meier, Elizabeth L;Daitch, Allison K;Yao, Qing;Bhargava, Anant;Jensen, Grant J;Goley, Erin D
• 通讯作者: Goley, Erin D

Programmed Secretion Arrest and Receptor-Triggered Toxin Export during Antibacterial Contact-Dependent Growth Inhibition.

抗菌接触依赖性生长抑制过程中的程序性分泌抑制和受体触发的毒素输出。

• 发表时间: 2018
• 影响因子: 64.5
• 作者: Ruhe, Zachary C;Subramanian, Poorna;Song, Kiho;Nguyen, Josephine Y;Stevens, Taylor A;Low, David A;Jensen, Grant J;Hayes, Christopher S
• 通讯作者: Hayes, Christopher S

Structure of the Bacterial Cellulose Ribbon and Its Assembly-Guiding Cytoskeleton by Electron Cryotomography.

通过电子冷冻断层扫描技术研究细菌纤维素带的结构及其组装引导细胞骨架。

• 发表时间: 2021
• 影响因子: 3.2
• 作者: Nicolas, William J;Ghosal, Debnath;Tocheva, Elitza I;Meyerowitz, Elliot M;Jensen, Grant J
• 通讯作者: Jensen, Grant J

Assigning chemoreceptors to chemosensory pathways in Pseudomonas aeruginosa.

将化学感受器分配给铜绿假单胞菌的化学感应途径。

• 发表时间: 2017-11-28
• 影响因子: 11.1
• 作者: Ortega, Davi R;Fleetwood, Aaron D;Krell, Tino;Harwood, Caroline S;Jensen, Grant J;Zhulin, Igor B
• 通讯作者: Zhulin, Igor B