CAREER: Organization and Force Production in Bacteria

在这个CAREER提案中，PI提议研究生物学中的一个基本问题：纳米尺度的蛋白质如何在细胞水平上产生微米尺度的力？具体来说，PI将确定细菌中染色体分离和细胞分裂的机制。拟议的实验将通过(i)成像，（ii）机械扰动和（iii）力测量来研究细菌的细胞内组织和力产生。超分辨率，三维成像将用于量化细胞内细胞骨架蛋白的几何构象以及它们如何响应外部施加的力。主动分离的DNA区域的三维轨迹将揭示使用单粒子跟踪。此外，局部细胞曲率在细胞分裂平面位置中的作用以及在细胞分裂过程中收缩环产生机械力的能力将用原子力显微镜进行探测。拟议的教育计划通过(i)课程开发，（ii）动手观察和（iii）多媒体细胞环境为未来的生物物理学家提供学习机会。通过他在普林斯顿大学物理系和Lewis-Sigler整合基因组学研究所的联合任命，PI将开发和教授两门关于生物物理学的新课程，将研究成果整合到更广泛的生物物理学背景中。为了激励下一代科学家，PI将与当地中学生合作，作为普林斯顿大学科学与工程博览会的一部分，通过显微镜和建模活动向他们介绍细胞运动的物理原理。一个交互式软件程序，部分基于该提案的成像数据，将允许学生探索存在于细胞内的拥挤而迷人的三维世界。该提案的长期目标是开发新的方法来研究生命系统中的单分子动力学，并了解不同的长度和时间尺度是如何整合为一个活的细菌或细胞出现的。这个项目涵盖了细胞生物学、分子生物物理学、物理学和计算生物学的各个方面。参与该项目的学生将在几个科学领域的交叉领域接受培训。拟议的研究具有真正具有变革性的所有要素。该项目由物理科学理事会物理部的生命系统物理项目分子和生物科学理事会的分子和细胞生物科学部共同支持。

In this CAREER proposal the PI proposes to study a fundamental question in biology: How do nanometer-scale proteins give rise to micron-scale forces at the cellular level? Specifically, the PI will determine the mechanisms of chromosome segregation and cell division in bacteria. The proposed experiments will investigate intracellular organization and force generation in bacteria through (i) imaging, (ii) mechanical perturbation and (iii) force measurement. Super-resolution, three-dimensional imaging will be used to quantify the geometric conformations of cytoskeletal proteins inside cells and how they respond to externally applied forces. The three-dimensional trajectories of actively segregated regions of DNA will be revealed using single particle tracking. In addition, the role of local cell curvature in the placement of the cell division plane and the ability of the contractile ring to generate mechanical force during cell division will be probed with atomic force microscopy. The proposed education plan provides learning opportunities for future biophysical scientists through combined (i) course development, (ii) hands-on observations and (iii) a multimedia cellular environment. The PI will develop and teach two new courses on Biological Physics that integrate research results in a broader biophysical context through his joint appointment in the Department of Physics and the Lewis-Sigler Institute for Integrative Genomics at Princeton University. To motivate the next generation of scientists, the PI will work with local middle school students as part of Princeton Universitys Science and Engineering Expo, introducing them firsthand to the physics of cellular locomotion through microscopy and modeling activities. An interactive software program, partly based on imaging data from this proposal, will be created that allows students to explore the crowded and fascinating three-dimensional world that exists inside cells.The long term goal of the proposal is to develop novel methods for studying single molecule dynamics in living systems and to understand how different length and time scales are integrated for a living bacteria or cell to emerge. This project encompasses aspects of cell biology, molecular biophysics, physics, and computational biology. The students working on the project will be trained at the interface of several fields of science. The proposed research has all ingredients to be truly transformational. This project is jointly supported by the Physics of Living Systems Program Molecular in the Division of Physics at the Physical Sciences Directorate and by the the Division of Molecular and Cellular Biosciences in the Directorate for Biological Sciences.