Cell Wall Formation in Rod Shaped Bacteria

杆状细菌细胞壁的形成

• 批准号: BB/Y003187/1
• 负责人: David Ian Roper
• 金额: $ 430.86万
• 依托单位: University of Warwick
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2024
• 资助国家: 英国
• 起止时间: 2024 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FY003187%2F1
• 关键词: Cell; Wall; Formation; Rod; Shaped

PURPOSE OF RESEARCHBacteria are surrounded by a mesh-like cell wall, made out of a polymer called peptidoglycan, which gives them their shape and protects them from bursting due to high internal turgor pressure. In many rod-shaped bacteria, including pathogens such as Escherichia coli, Salmonella species and Pseudomonas aeruginosa, cells grow by adding new material around the cell sidewall. This is principally performed by a multi-protein complex called the elongasome which maintains the cell's rod shape by precise insertion of new cell wall material. Inactivation of the elongasome leads to loss of rod-shape and severely impaired cell fitness. Antimicrobial resistance now represents a major threat to modern medicine, motivating the urgent search for fundamentally new ways to attack the formation of the bacterial cell wall. Elongasome proteins are structurally very similar to the divisome cell wall synthesis proteins that divide the cell, and together, elongasome and divisome proteins represent the primary architects of the bacterial cell wall. Therefore, the research into the molecular basis of elongasome function proposed will not only advance our understanding of how the bacterial cell wall is built but may also identify new routes to next generation antibiotics that target elongasome and divisome cell wall synthesis proteins and are effective against a wide range of bacteria.In this project we will determine the basis of elongasome function and regulation from an atomic to macromolecular scale. This will be achieved using synergy between advanced molecular imaging techniques, advanced biochemical and structural biology analysis, cutting-edge computational analysis and simulation methods and bespoke chemical probes, in a fundamentally integrative team-science research programme.TIMELINESS- We recently solved the structure of the E. coli RodA-PBP2 core of the elongasome, providing a foundational basis for understanding the molecular mechanism of elongasome function- We have developed a repertoire of new approaches to realise this project including new chemical biology tools based on elongasome substrate mimetics, innovative molecular simulations of multi-protein complexes and new methods for single molecule biophysics of cell wall synthesis proteins- This makes us uniquely placed to make major advances in our understanding of the elongasome and the molecular basis of bacterial cell wall synthesis.VALUE FOR MONEYThe highly integrated team science work packages proposed here will enable major advances in bacterial cell wall biology that are simply not possible in an ordinary responsive model proposal. Direct added value is provided by sustained strategic investment at Warwick to build a national centre of excellence in bacterial cell envelope biology, including:- Recruitment of Stansfeld (2019) and Holden (2022) - £54M investment in the new laboratory complex occupied by all Warwick applicants, - A £1.7M charitable donation for a Howard Dalton research centre which provides underpinning support for cell wall research and funds a £120K smFRET instrument heavily used in this in this proposal. Team science is greatly facilitated by location of Warwick applicants in the same new building and established collaborations between all Warwick and Belfast applicants. OUTCOMES-Fundamental knowledge gain: This proposal approach will reveal broad insights into the molecular function and regulation of the SEDS-PBP proteins that are the major architects of the bacterial cell wall. -Potential applications in biomedicine and biotechnology: This will lay the foundations for future antimicrobial therapies targeting cell wall synthesis by SEDS proteins. -Staff training: We will train a cohort of interdisciplinary pre and postdoctoral scientists orientated in an area of microbiology that desperately needs new talent and enable future generations of antimicrobial research in academia and industry.

研究目的细菌被一层网状细胞壁包围着，这种细胞壁是由一种叫做肽聚糖的聚合物构成的，这种聚合物赋予了细菌形状，并保护它们不因内部高膨胀压力而破裂。在许多杆状细菌中，包括大肠杆菌、沙门氏菌和铜绿假单胞菌等病原体，细胞通过在细胞侧壁周围添加新物质来生长。这主要是由一种称为长体的多蛋白复合物完成的，它通过精确插入新的细胞壁材料来保持细胞的棒状。长体的失活导致杆状体的丧失和细胞适应性的严重受损。抗菌素耐药性现在是对现代医学的主要威胁，促使人们迫切寻求从根本上攻击细菌细胞壁形成的新方法。细长体蛋白在结构上与分裂体细胞壁合成蛋白非常相似，它们共同构成了细菌细胞壁的主要结构。因此，对长体功能的分子基础的研究不仅将促进我们对细菌细胞壁如何构建的理解，而且还可能确定针对长体和分裂体细胞壁合成蛋白的下一代抗生素的新途径，并对多种细菌有效。在这个项目中，我们将确定从原子到大分子尺度的细长体功能和调控的基础。这将通过先进的分子成像技术、先进的生物化学和结构生物学分析、尖端的计算分析和模拟方法以及定制的化学探针之间的协同作用，在一个基本整合的团队科学研究计划中实现。及时性-我们最近解决了大肠杆菌长体RodA-PBP2核心的结构，为理解长体功能的分子机制提供了基础基础-我们已经开发了一系列新方法来实现这个项目，包括基于长体底物模拟物的新化学生物学工具，多蛋白复合物的创新分子模拟和细胞壁合成蛋白的单分子生物物理学新方法-这使我们在理解长体和细菌细胞壁合成的分子基础方面取得了重大进展。这里提出的高度集成的团队科学工作包将使细菌细胞壁生物学取得重大进展，这在普通的响应模型建议中根本不可能实现。华威通过持续的战略投资提供直接附加值，以建立一个国家细菌包膜生物学卓越中心，包括：-斯坦斯菲尔德（2019年）和霍尔顿（2022年）的招聘-为所有华威申请人所占据的新实验室综合体投资5400万英镑，-为霍华德道尔顿研究中心提供170万英镑的慈善捐赠，该中心为细胞壁研究提供基础支持，并资助12万英镑的smFRET仪器在本提案中大量使用。华威申请人在同一栋新大楼的位置以及所有华威和贝尔法斯特申请人之间建立的合作关系极大地促进了团队科学。结果：基础知识的获得：这一提议的方法将揭示对细菌细胞壁的主要构建者SEDS-PBP蛋白的分子功能和调控的广泛见解。-在生物医学和生物技术方面的潜在应用：这将为未来针对SEDS蛋白合成细胞壁的抗菌治疗奠定基础。-人员培训：我们将在微生物学领域培养一批跨学科的博士后和博士后科学家，该领域迫切需要新的人才，并使学术界和工业界的未来几代抗菌素研究成为可能。