cdiGMP regulation of Biofilm Formation

在接下来的几年里，我们的目标是了解运动微生物，如铜绿假单胞菌，是如何调节细胞的生长的。 过渡到生物膜生活方式。这种转变的一个关键方面是监管（也许“调制”是一个 这种微生物的三种表面行为：鞭毛介导的群集， 抽搐运动和EPS依赖的生物膜形成。下面提出的实验试图 理解这些表面行为协调的机制。的一个组成部分 这项工作正在定义这种微生物如何检测表面接触;我们对PilY 1的研究使我们能够 剖析一个这样的“表面感应”途径。我的团队已经在这个令人兴奋的领域做出了贡献 在下一个资助期内，我们会继续推行这方面的工作，包括：（i）完成 在最初的申请中提出的，（ii）打破了新的研究领域。为了解决后者，我 我建议将一个新的正式的和资助的合作纳入6-10年的项目与博士杰拉德 加州大学洛杉矶分校的王。我们已经共同发表了五篇论文，第六篇论文在PNAS出版。我们提出 以下研究继续检验中心假设，即cdG水平在 对细胞与基质接触的反应，导致生物膜的运动性降低和促进 阵 目标1。检验蛋白质-蛋白质相互作用级联信号从PilY 1到SadC， 调节c-di-GMP水平。 目标2.检验SadC通过内分泌途径协调群集和抽搐运动的假设。 膜复合物 目标3。测试定子交换在早期依恋中驱动不同行为的假设。 目标4。检验鞭毛和TFP介导的运动调节有助于 多微生物群落的形成。 目标5。检验c-di-GMP信号传导是维持成熟生物膜所必需的假设。 相关性（参见说明）： 据估计，基于细菌生物膜的感染每年造成超过10亿美元的增加。 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， 从而提供了如何预防此类感染的见解。

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.