Probing the molecular interactions and architecture of bacterial cell division proteins

探究细菌细胞分裂蛋白的分子相互作用和结构

• 批准号: RGPIN-2015-06104
• 负责人: Khursigara, Cezar
• 金额: $ 2.48万
• 依托单位: University of Guelph
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=655490
• 关键词: Probing; molecular; interactions; architecture; bacterial

Cellular microbiology has seen a renaissance in the past several years, spurred in part by advances in imaging techniques. Our ability to peer into cells to see the localization and organization of macromolecular complexes involved in fundamental processes has advanced our understanding of how bacteria move, associate and divide. In gram-negative bacteria, a group of proteins known as the divisome drives the synchronized invagination of the inner and outer membranes and peptidoglycan synthesis at the newly formed septum. Divisome proteins form the multicomponent Z-ring structure, which is primarily composed of the tubulin homologue FtsZ. The dynamics of FtsZ filament formation are considered essential for proper Z-ring assembly. In Escherichia coli, ~10 essential and over 20 nonessential proteins are involved in divisome assembly. Interactions occur in a hierarchical manner, and the proposed mechanisms of cell division can be divided into three distinct stages - early, intermediate and late. Although bacterial cell division has been intensively studied at the cellular and molecular levels for decades, with many of the key protein players identified, critical questions remain about the molecular mechanisms and protein interactions that govern the formation and dynamics of the divisome. ***This research program aims to elucidate the protein interactions and macromolecular structures involved in Z-ring stability and dynamics. Specifically, we will identify and characterize the interactions and macromolecular architecture of FtsZ and the Zap family of proteins. These interactions are critical for early-stage cell division, and the overlapping functions of the Zap proteins have been implicated in Z-ring-mediated septation. We will also determine the protein interactions in intermediate-stage cell division that govern how FtsK, an essential cell division `checkpoint' protein, links division of the cytoplasm with septation and cell wall invagination. We will address these questions using an original combination of biochemical, molecular and cutting-edge imaging tools, which will lead to groundbreaking advances in cell division. The long-term vision for this research program is to develop a complete biochemical and structural picture of bacterial cell division during all of its distinct stages, thereby elucidating the mechanisms that drive this essential process. Our findings and the approaches we develop will transcend prokaryotic cell biology and will be applicable to diverse biological systems.**

在过去的几年中，细胞微生物学已经获得了复兴，部分原因是成像技术的进步。我们凝视细胞的能力，看到与基本过程中涉及的大分子络合物的定位和组织有关，这使我们对细菌如何移动，伴侣和分裂进行了理解。在革兰氏阴性细菌中，一组称为Divisome的蛋白质驱动了内部和外膜的同步内部，以及新形成的隔膜的PepperyDoglycan合成。除了多组分Z环结构，该蛋白主要由微管蛋白同源FTSZ组成。 FTSZ丝形成的动力学对于适当的Z形组件被认为是必不可少的。在大肠杆菌中，〜10个必需的和20多种非必要蛋白参与了分裂组装。相互作用以层次的方式发生，细胞分裂的提议机制可以分为三个不同的阶段 - 早期，中间和晚期。尽管细菌细胞分裂数十年来一直在细胞和分子水平上进行了大量研究，但由于许多关键蛋白质参与者都确定了许多关键蛋白质，但有关分子机制和蛋白质相互作用的关键问题仍然是控制了分裂体的形成和动力学。 ***该研究计划旨在阐明参与Z形环稳定性和动力学的蛋白质相互作用和大分子结构。具体而言，我们将识别并表征FTSZ和ZAP蛋白质家族的相互作用和大分子结构。这些相互作用对于早期细胞分裂至关重要，ZAP蛋白的重叠功能已在Z-RING介导的分隔中实现。我们还将确定中间阶段细胞分裂中的蛋白质相互作用，该蛋白质是如何控制细胞分裂的“检查点”蛋白的FTSK，将细胞质的分裂与分离和细胞壁的内陷联系起来。我们将使用生化，分子和尖端成像工具的原始组合来解决这些问题，这将导致细胞分裂的突破性进步。该研究计划的长期视野是在其所有不同阶段中开发出细菌细胞分裂的完整生化和结构图，从而阐明了推动这一基本过程的机制。我们的发现和我们开发的方法将超越原核细胞生物学，并适用于潜水生物系统。**