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 我实验室的研究旨在解决内部存在哪些大型大分子复合物的问题 单元格以及其功能如何取决于它们的结构和位置。为了实现这一目标，我们使用 电子冷冻理学（ECT）以图像完整细胞内3D中的天然细胞结构，执行 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 艾滋病毒和冷冻EM方法的开发。艾滋病毒的工作与我们的其他项目相辅相成 驱动器并受同一技术开发的驱动，但它是单独资助的。在微生物细胞生物学中， 在接下来的五年中，我们旨在提高我们对：（1） 细菌分泌系统； （2）在生命的所有三个王国中，细胞分裂的机制； （3） 细菌细胞骨架和DNA分离丝。驾驶这些项目，以冷冻EM方法 开发，我们的目标是：（1）扩展ECT的吞吐量和自动化； （2）应用低温浓缩离子 横梁开孔以将ECT的覆盖范围扩展到任何细胞，而不论大小如何； （3）使用我们的ECT测量 告知实际的粗粒生物过程的粗粒元模型以开发和测试 机械假设。

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

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

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

A bacterial membrane sculpting protein with BAR domain-like activity.

• DOI: 10.7554/elife.60049
• 发表时间: 2021-10-13
• 期刊: eLife
• 影响因子: 7.7
• 作者: Phillips DA;Zacharoff LA;Hampton CM;Chong GW;Malanoski AP;Metskas LA;Xu S;Bird LJ;Eddie BJ;Miklos AE;Jensen GJ;Drummy LF;El-Naggar MY;Glaven SM
• 通讯作者: Glaven SM