RpoS Regulation of Borrelia burgdorferi Genes in vivo

伯氏疏螺旋体基因的体内 RpoS 调控

基本信息

批准号：
10459323
负责人：
MELISSA J CAIMANO
金额：
$ 59.38万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
1990
资助国家：
美国
起止时间：
1990-04-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10459323
关键词：
Address Adopted Arbovirus Infections Area Arthropod Vectors Arthropods Back Bacteria Bacteria sigma factor KatF protein Binding Proteins Borrelia Borrelia burgdorferi Centers for Disease Control and Prevention (U.S.)Complex Cues DNA-Directed RNA Polymerase Data Dialysis procedure Environment Gatekeeping Gene Expression Regulation Genes Genetic Genetic Transcription Holoenzymes Human Individual Infection Ixodes Ixodidae Laboratories Life Style Lyme Disease Mammals Mediating Mediator of activation protein Membrane Methodology Mission Modeling Mus Mutagenesis Nature Order Spirochaetales OspA protein Output Pathway interactions Periodicity Phase Proteome Publications Regulation Regulatory Pathway Regulon Reporting Repression Rodent Role Signal Transduction Signaling Molecule Testing Thinking Ticks Time United States Virulence Work antagonist cohort enzootic experimental study gene function holistic approach in vivo metabolome programs promoter response tick transmission transcriptome transcriptomics vector tick

项目摘要

Borrelia burgdorferi (Bb), the agent of Lyme disease (LD), is maintained in nature within a complex enzootic cycle involving a mammalian reservoir host and a tick vector. To sustain this cycle, Bb must adjust its transcriptome, proteome, and metabolome to arthropod- and mammalian host-derived signals as it shuttles between the two. The Rrp2/RpoN/RpoS pathway has gained widespread recognition as a central player in borrelial gene regulation. Our laboratory has long been at the forefront of efforts to characterize the cohort of genes controlled by RpoS in both the tick and mammal. Unfortunately, we still do not know the entire output of RpoS-RNA polymerase (RNAP) holoenzyme in nature. The present proposal addresses this shortfall by combining state-of-the-art transcriptomic and mutagenesis methodologies to determine where/when individual genes contribute to RpoS's dual-host `mission'. In prior publications, we were instrumental in delineating the temporal boundaries of the RpoS-ON and –OFF states in vivo. In recent years, however, our thinking about this dichotomy has become much more nuanced. We now believe that the contours of the RpoS regulon change profoundly and in host-specific fashion during the RpoS-ON state These results, in concert with studies of the Hk1/Rrp1 pathway, give rise to our central hypothesis--the dynamic nature of the RpoS regulon reflects the confluence of non-RpoS regulatory pathways with mechanisms that govern the ON/OFF states of RpoS and the output of RpoS-RNAP. Along these lines, we now propose that the tick-phase signaling molecule c-di-GMP is crucial to this regulatory cross-talk. Moreover, we and others have found that the c-di-GMP-binding protein PlzA promotes expression of RpoS and, hence, Bb virulence in the mouse, the stage of the enzootic cycle in which the Hk1/Rrp1 pathway is OFF. Our efforts to clarify this bifunctional role of PlzA as a mediator of c-di-GMP signaling in ticks and a c-di-GMP-independent regulator of RpoS in mice takes our field into uncharted territory. Lastly, in years past, our analysis of mammalian host-adapted spirochetes cultivated in dialysis membrane chambers revealed that RpoS not only upregulates genes required for tick transmission and mammalian infection but also represses genes required for colonization and adaptation to the tick. Our recent work suggests that RpoS-RNAP directly represses transcription of tick-phase genes by occluding their σ70 promoters. We will test this `RpoS as repressor' model and explore new data that c-di-GMP antagonizes RpoS-mediated repression in mammalian host-adapted Bb. Our long-term objective is to achieve an integrative understanding of how the Rrp2/RpoN/RpoS pathway fulfills its essential mission--guiding LD spirochetes from tick to mouse and back again. We will accomplish this by defining the contours of the RpoS regulon in ticks and mice (Aim 1); clarifying the convergence of c-di-GMP- and PlzA-dependent signaling with the RpoS pathway (Aim 2); and dissecting RpoS-mediated repression of tick-phase genes and its antagonism by c-di-GMP in mammalian host-adapted Bb (Aim 3).

莱姆病（LD）的药物（LD）的伯氏伯氏菌（BB）在复杂的enzootic中维持在自然界中循环涉及哺乳动物的储藏室和一个tick矢量。为了维持这个周期，BB必须调整其转录组，蛋白质组和代谢组，乘坐乘坐宿主的信号在两个之间。 RRP2/RPON/RPOS途径已获得宽度的认可，作为中心参与者疏松基因调节。我们的实验室长期以来一直处于表征的努力中 tick和哺乳动物中RPO控制的基因。不幸的是，我们仍然不知道自然界中的RPOS-RNA聚合酶（RNAP）全酶。本提案通过结合最新的转录组和诱变方法，以确定单个何时/何时基因促进了RPO的双主机“任务”。在先前的出版物中，我们有助于描述 RPOS-on和 - off状态在体内的临时边界。但是，近年来，我们对此的思考二分法变得更加细微。我们现在相信RPOS Regulon的轮廓变化在RPOS-ON状态期间，以特定于宿主为特定的方式这些结果，与研究有关 HK1/RRP1途径，引起我们的中心假设 - RPOS常规的动态性质反映了非RPOS调节途径的汇合与控制RPO的ON/OFF状态的机制和 RPOS-RNAP的输出。沿着这些线，我们现在建议tick相信号分子C-DI-GMP为对于此调节性串扰至关重要。此外，我们和其他人发现C-DI-GMP结合蛋白PLZA 促进RPO的表达，因此，小鼠中的BB病毒是Enzootic循环的阶段，其中 HK1/RRP1途径已关闭。我们为阐明PLZA作为C-DI-GMP信号传导的中介者的双功能作用的努力在小鼠中，RPO的tick和C-DI-GMP的调节器中，我们的领域进入了未知的领域。最后，在过去的几年中，我们对在透析膜室中种植的哺乳动物宿主适应的螺旋体的分析发现RPO不仅上调了tick传播和哺乳动物感染所需的基因压抑定植所需的基因并适应壁虱。我们最近的工作表明RPOS-RNAP 直接通过遮挡tick相基因的转录来反映其σ70启动子的转录。我们将测试此`rpos 阻遏物的模型并探索C-DI-GMP拮抗RPOS介导的表示的新数据主机适应的BB。我们的长期目标是对如何融合了解 RRP2/RPON/RPOS途径实现了其必不可少的任务 - 指导LD Spirochetes从tick到鼠标和背部再次。我们将通过定义tick和小鼠中RPOS Regulon的轮廓来实现这一目标（AIM 1）；澄清 C-DI-GMP和PLZA依赖性信号与RPOS途径的收敛性（AIM 2）；并解剖 RPOS介导的tick相基因的表示及其对哺乳动物宿主适应BB中C-DI-GMP的拮抗作用（目标3）。

项目成果

期刊论文数量（47）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Of ticks, mice and men: understanding the dual-host lifestyle of Lyme disease spirochaetes.

DOI：
10.1038/nrmicro2714
发表时间：
2012-01-09
期刊：
Nature reviews. Microbiology
影响因子：
0
作者：
通讯作者：

BosR and PlzA reciprocally regulate RpoS function to sustain Borrelia burgdorferi in ticks and mammals.

DOI：
10.1172/jci166710
发表时间：
2023-03-01
期刊：
JOURNAL OF CLINICAL INVESTIGATION
影响因子：
15.9
作者：
Grassmann, Andre A.;Tokarz, Rafal;Golino, Caroline;McLain, Melissa A.;Groshong, Ashley M.;Radolf, Justin D.;Caimano, Melissa J.
通讯作者：
Caimano, Melissa J.

The FUR-like regulators PerRA and PerRB integrate a complex regulatory network that promotes mammalian host-adaptation and virulence of Leptospira interrogans.