Mechanisms and Regulation of Cell Division in Bacteria

细菌细胞分裂的机制和调控

• 批准号: 10373994
• 负责人: WILLIAM MARGOLIN
• 金额: $ 42.76万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10373994
• 关键词: Actins; Address; Anabolism; Antibiotics; Bacteria; Bacterial Model; Biomimetics; Biophysics; Bypass; Cell Wall; Cell division; Cell physiology; Cells; Cities; Collaborations; Cues; Cytokinesis; Cytology; Escherichia coli; Genetic Materials; Growth; Image; In Vitro; Infrastructure; Investigation; Maps; Membrane Proteins; Microscopy; Molecular Genetics; Polymers; Proliferating; Protein Biochemistry; Proteins; Public Health; Regulation; Resistance; Resolution; Role; Structure; System; Time; Tubulin; Work; cell growth; flexibility; genetic approach; insight; medical specialties; nanomachine; new therapeutic target; novel; reconstitution; response; success

Project summary A cell is like a city, with an organized yet dynamic infrastructure grouped into specialties. For the last 25 years, my lab has investigated how the simplest cells—bacteria—organize themselves and divide to make progeny cells. We mainly focus on how bacteria such as E. coli achieve the daunting task of splitting themselves in two at just the right time (once their genetic material is duplicated) and place (exactly in the middle) every 20 minutes without making errors. The keys to this success are ancient and universal versions of protein polymers of actin (FtsA) and tubulin (FtsZ), which our lab visualized for the first time in living bacteria over 20 years ago. Today, we use state of the art super-resolution imaging, combined with molecular genetics, protein biochemistry, interaction studies, and in vitro reconstitution, to gain more detailed insights into the structure and regulation of these cytoskeletal polymers and their associated proteins, which comprise the dynamic membrane-associated protein nanomachine (divisome) that divides bacterial cells. Thanks in part to our characterization of bypass suppressors of essential divisome proteins, it is now becoming clear that the divisome is highly flexible, and can remodel itself in response to various inputs and perturbations. Despite impressive contributions by many labs, there is much to be learned about overall divisome structure, the interchangeability of its parts, and how it remodels in response to temporal and environmental cues. We will address these fundamental questions by (1) obtaining more high-resolution information about protein-protein contacts during cytokinesis by combining biophysical, cytological, and genetic approaches; (2) investigating the role of oligomeric state of FtsZ and FtsA in divisome function and regulation, using super-resolution microscopy of whole cells and reconstituted biomimetic protein-membrane systems; (3) taking advantage of the diversity of divisome proteins from other model bacterial species to distinguish between common and specialized mechanisms; (4) understanding the interplay between the divisome and other large-scale cellular processes such as cell wall biosynthesis. We will leverage these approaches by continuing our collaborations with several close colleagues who have complementary interdisciplinary expertise. Our ongoing investigation of how the simplest cells divide should pave the way for an unprecedented understanding of how an entire cell functions and reproduces. Having an accurate map of that city-cell's dynamic infrastructure will allow predictions to be made about how it works, and how to disrupt it.

项目概要 细胞就像一座城市，拥有按专业划分的有组织且动态的基础设施。 过去 25 年，我的实验室研究了最简单的细胞（细菌）如何自我组织和 我们主要关注大肠杆菌等细菌如何实现这一点。 在适当的时间将自己一分为二（一旦它们的遗传物质被分离出来）是一项艰巨的任务 重复）并每 20 分钟放置一次（正好在中间），不要出错。 这一成功是肌动蛋白 (FtsA) 和微管蛋白蛋白质聚合物的古老且通用的版本 (FtsZ)，我们的实验室在 20 多年前首次在活细菌中观察到它，今天我们使用它。 最先进的超分辨率成像，结合分子遗传学、蛋白质生物化学、 相互作用研究和体外重建，以获得对结构和结构的更详细的了解 这些细胞骨架聚合物及其相关蛋白质的调节，其中包括 分裂细菌细胞的动态膜相关蛋白纳米机器（分裂体）。 部分归功于我们对必需分裂体蛋白旁路抑制子的表征，它是 现在越来越清楚的是，分裂体是高度灵活的，并且可以根据 尽管许多实验室做出了令人印象深刻的贡献，但仍有很多工作要做。 了解整体的分割结构、其部件的互换性以及它如何改造 我们将通过以下方式解决这些基本问题： (1) 通过以下方式获得有关胞质分裂过程中蛋白质-蛋白质接触的更高分辨率信息 结合生物物理、细胞学和遗传学方法；（2）研究寡聚体的作用； 使用超分辨率显微镜观察 FtsZ 和 FtsA 在分裂功能和调节中的状态 全细胞和重建仿生蛋白膜系统；（3）利用 来自其他模型细菌物种的分裂蛋白的多样性，以区分常见的细菌 和专门机制；（4）理解分裂与其他之间的相互作用 我们将利用这些方法来进行大规模的细胞过程，例如细胞壁生物合成。 继续与几位具有互补性的密切同事合作 跨学科的专业知识。 我们正在进行的关于最简单的细胞如何分裂的研究应该为 对整个细胞如何运作和繁殖有着前所未有的准确了解。 该城市单元动态基础设施的地图将允许对其工作方式进行预测， 以及如何破坏它。