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.

项目摘要 我的实验室的研究试图解决内部存在哪些大的大分子复合体的问题 以及它们的功能是如何由它们的结构和位置决定的。为了实现这一目标，我们使用 电子冷冻断层扫描(ECT)在完整细胞内对自然细胞结构进行3D成像，进行 活体结构生物学。该实验室的项目分为三类：微生物细胞生物学、结构生物学 以及冷冻-EM方法的开发。艾滋病毒工作与我们的其他项目相辅相成，因为它 驱动并由相同的技术开发驱动，但它是单独资助的。在微生物细胞生物学中， 在接下来的五年里，我们的目标是提高我们对以下方面的理解：(1) 细菌分泌系统；(2)所有三个生命王国的细胞分裂机制；以及(3) 细菌细胞骨架和DNA分离细丝。以低温电磁方法推动这些项目 发展，我们的目标是：(1)扩大ECT的产量和自动化；(2)应用低温聚焦离子 将ECT的覆盖范围扩展到任何单元，而不考虑大小；以及(3)使用我们的ECT测量来 通知开发生物过程的实际粗粒度模型以进行开发和测试 机械论假说。

项目成果

Challenges in solving structures from radiation-damaged tomograms of protein nanocrystals assessed by simulation.

• DOI: 10.1107/s2059798321002369
• 发表时间: 2021-05-01
• 期刊: Acta crystallographica. Section D, Structural biology
• 作者: Peck A;Yao Q;Brewster AS;Zwart PH;Heumann JM;Sauter NK;Jensen GJ
• 通讯作者: Jensen GJ

Structure of the fission yeast actomyosin ring during constriction.

收缩过程中裂变酵母肌动球蛋白环的结构。

• DOI: 10.1073/pnas.1711218115
• 发表时间: 2018
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Swulius,MatthewT;Nguyen,LamT;Ladinsky,MarkS;Ortega,DaviR;Aich,Samya;Mishra,Mithilesh;Jensen,GrantJ
• 通讯作者: Jensen,GrantJ

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

• DOI: 10.1016/j.cell.2018.10.033
• 发表时间: 2018-11-01
• 期刊: Cell
• 影响因子: 64.5
• 作者: Ruhe ZC;Subramanian P;Song K;Nguyen JY;Stevens TA;Low DA;Jensen GJ;Hayes CS
• 通讯作者: Hayes CS

Cryo-electron tomography of the onion cell wall shows bimodally oriented cellulose fibers and reticulated homogalacturonan networks.

• DOI: 10.1016/j.cub.2022.04.024
• 发表时间: 2022-06-06
• 期刊: CURRENT BIOLOGY
• 影响因子: 9.2
• 作者: Nicolas, William J.;Faessler, Florian;Dutka, Przemyslaw;Schur, Florian K. M.;Jensen, Grant;Meyerowitz, Elliot
• 通讯作者: Meyerowitz, Elliot

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

• DOI: 10.1128/jb.00371-20
• 发表时间: 2021-01-11
• 期刊: Journal of bacteriology
• 影响因子: 3.2
• 作者: Nicolas WJ;Ghosal D;Tocheva EI;Meyerowitz EM;Jensen GJ
• 通讯作者: Jensen GJ