Mapping the Interactions and Dynamics that Organize Bacteria Cells

项目摘要 分子规模的相互作用使亚细胞组织成为可能，但目前最先进的技术还不包括 测量分子在纳米尺度上和实际中如何运动和协作的通用方法 单元格内的时间。这一差距在细菌细胞中尤为明显。因此，我们对本组织的描述 细菌细胞的部分是不完整的。本提案旨在了解蜂窝组件如何组织-by 脚手架、聚集、相分离或其他--以产生细菌细胞的一般模型 组织，并最终使我们能够促进共生细菌或抗击人类疾病。因此，我们的目标是 测量活细菌细胞中不同蛋白质分子的位置、相互作用和运动 与这些细胞的亚细胞组织有关的系统。这项提案将在下一步发展这些- 通过三个协同的具体目标生成超分辨率工具：(1)超分辨率如何单一- 活细菌细胞中分子动力学在空间和时间上的变化；(2)超分辨亚细胞相互作用 在活细菌细胞的空间和时间上；以及(3)改进活细菌中单分子的检测 细胞。我们开发的绘制分子运动和相互作用图的工具包将广泛适用于 单分子细菌学群落。此外，通过专注于细菌细胞生物学的应用， 该项目中开发的工具将通过使长期可用的 术语，微生物生物学中的重大问题。

Project Summary Molecular-scale interactions enable subcellular organization, but the current state-of-the-art does not include a generalizable method for measuring how molecules move and cooperate on the nanometer scale and in real time within the cell. This gap is particularly striking in bacteria cells. Accordingly, our picture of the organization of the bacterial cell is incomplete. This proposal aims to understand how cellular components organize—by scaffolding, aggregation, phase separation, or otherwise—to produce a general model of bacterial cell organization and ultimately enable us to promote commensal bacteria or fight human disease. Thus, we aim to measure the positioning, interactions, and motions of molecules in living bacterial cells in different protein systems implicated in the sub-cellular organization of these cells. This proposal will develop these next- generation super-resolution tools through three synergistic specific aims: (1) to super-resolve how single- molecule dynamics vary in space and in time in living bacterial cells; (2) to super-resolve sub-cellular interactions in space and time in living bacteria cells; and (3) to improve the detection of single molecules in living bacteria cells. The toolkit that we develop to map molecular motions and interactions will be broadly applicable to the single-molecule bacteriology community. Furthermore, by focusing on applications in bacterial cell biology, the tools developed in this project will have a widespread, positive biomedical impact by making accessible long- term, significant questions in microbial biology.