cdiGMP regulation of Biofilm Formation

生物膜形成的 cdiGMP 调节

• 批准号: 10219049
• 负责人: George A. O'Toole
• 金额: $ 52.2万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10219049
• 关键词: Address; Area; Behavior; Cells; Collaborations; Communities; Complex; Funding; Goals; Infection; Instruction; Life Style; Mediating; Medical Care Costs; Membrane; Microbe; Microbial Biofilms; Microbiology; Paper; Pathway interactions; Pseudomonas aeruginosa; Publishing; Regulation; Research; Signal Transduction; Surface; Testing; Work; base; cell motility; experimental study; insight; prevent; protein protein interaction; response

In the coming years, our goal is to understand how motile microbes, such as P. aeruginosa, regulate the transition to a biofilm lifestyle. A key aspect of this transition is the regulation (perhaps “modulation” is a better word) of the three surface behaviors of this microbe: flagellar mediated swarming, TFP-mediated twitching motility and EPS-dependent biofilm formation. The experiments proposed below seek to understand the mechanism(s) underlying the coordination of these surface behaviors. An integral aspect of this work is defining how this microbe detects surface engagement; our work on PilY1 is allowing us to dissect one such “surface sensing” pathway. My group has already made contributions to this exciting area of microbiology, and we will continue to do so in the come funding period by (i) completing studies proposed in the original application and (ii) breaking open new areas of research. To address the latter, I propose to incorporate a new formal and funded collaboration into years 6–10 of the project with Dr. Gerard Wong at UCLA. We have already co-published five papers with a 6th paper in press at PNAS. We propose the following studies to continue to test the central hypothesis that cdG levels are up-regulated in response to cell-to-substratum contact, leading to reduced motility and promotion of biofilm formation. Aim 1. Test the hypothesis that a protein-protein interaction cascade signals from PilY1 to SadC to modulate c-di-GMP levels. Aim 2. Test the hypothesis that SadC coordinates swarming and twitching motility via inner membrane complexes. Aim 3. Test the hypothesis that stator swapping drives distinct behaviors during early attachment. Aim 4. Test the hypothesis that modulation of flagellar- and TFP-mediated motility contributes to the formation of polymicrobial communities. Aim 5. Test the hypothesis that c-di-GMP signaling is required to maintain mature biofilms. RELEVANCE (See instructions): Bacterial biofilm-based infections are estimated to cause upwards of a billion dollars annually in increased medical costs. The work we propose here will reveal mechanisms whereby these communities form, and thus provide insight into how to prevent such infections.

在接下来的几年里，我们的目标是了解活动微生物，如铜绿假单胞菌，是如何调节 过渡到生物膜生活方式。这一转变的一个关键方面是监管(或许“调制”是一种 更确切地说，这种微生物的三种表面行为：鞭毛介导的聚集、TFP介导的 抽动运动和EPS依赖的生物膜的形成。下面提出的实验旨在 理解这些表面行为协调背后的机制(S)。的一个不可分割的方面 这项工作定义了这种微生物如何检测表面接触；我们在PilY1上的工作使我们能够 剖析一条这样的“表面感应”途径。我的团队已经为这一令人兴奋的领域做出了贡献 我们将在未来的资助期间继续这样做，方法是：(I)完成研究 提出在原创应用和(Ii)开拓新的研究领域。为了解决后者，我 建议将新的正式和有资金支持的合作纳入项目的第6-10年，与Gerard博士 加州大学洛杉矶分校的王。我们已经在PNAS联合发表了五篇论文和第六篇论文。我们建议 以下研究将继续检验CDG水平上调的中心假说 对细胞与底物接触的反应，导致生物膜的运动性降低和促进 队形。 目的1.检验假设，蛋白质相互作用将信号从PilY1级联到SADC，从而 调节c-di-GMP水平。 目的2.验证SADC通过内侧协调群聚和抽动运动的假设 膜复合体。 目的3.检验定子交换在依恋早期驱动不同行为的假设。 目的4.验证鞭毛和TFP介导的运动调节有助于 多种微生物群落的形成。 目的5.验证c-di-GMP信号是维持成熟生物膜所必需的假设。 相关性(请参阅说明)： 据估计，以细菌生物膜为基础的感染每年造成超过10亿美元的损失 医疗费用。我们在这里提出的工作将揭示这些社区形成的机制，以及 从而为如何预防此类感染提供了洞察力。