Genomics of Gene Regulation in Porphyromonas gingivalis

描述（由申请人提供）：破译感染期间细菌和宿主之间的“对话”对于理解微生物发病机制和宿主防御至关重要。长期以来，牙周病一直被认为是细菌感染，实验室的长期目标是鉴定感染期间表达的牙龈卟啉单胞菌基因，即。细菌和宿主之间的交互作用的一个方面。生物体的遗传易处理性，加上完整的基因组序列和基因微阵列的可用性，使这一目标变得触手可及。细菌使用双组分信号转导系统来监测和应对环境的变化。在本申请中，我们测试了在牙龈卟啉单胞菌中这些系统响应于健康和疾病期间龈下裂隙中普遍存在的体内条件的假设。牙龈卟啉单胞菌的基因调控的分子机制，提出了一个全面的研究，这一生物体的新领域的研究。将分析反应调节子以鉴定它们控制的基因及其启动子靶点。我们将测试我们的假设，通过确定在牙周炎患者的龈下菌斑的体内环境中的一个子集的基因的表达。实验方法利用了我们在分子遗传学、全基因组转录谱、启动子靶定位和DNA序列分析方面的经验。目标是响应RFA，因为我们将分析牙龈卟啉单胞菌细胞内信号传导机制，协调对环境变化的反应。通过定义调控因子、它们控制的基因以及它们在体外和体内的表达模式，我们可以确定可能引发牙周炎的条件，并将合理的治疗干预靶向于调控分子开关。

DESCRIPTION (provided by applicant): Deciphering the "conversation" between bacterium and host during infection is central to understanding microbial pathogenesis and host defense. Periodontal diseases have long been recognized as bacterial infections, and a long-term goal of the laboratory is to identify P. gingivalis genes expressed during infection, ie. one side of the cross-talk between bacterium and host. The genetic tractability of the organism, combined with the availability of a completed genome sequence and gene microarrays, has made this goal approachable. Bacteria use two-component signal transduction systems to monitor and react to changes in their environment. In this application, we test the hypothesis that in P. gingivalis these systems are responsive to in vivo conditions prevailing in the subgingival crevice during health and disease. A comprehensive study is proposed of the molecular mechanisms of gene regulation in P. gingivalis, a new area of research with this organism. Response regulators will be analyzed to identify the genes they control, and their promoter targets. We will test our hypothesis by determining the expression of a subset of identified genes in the in vivo environment of subgingival plaque from periodontitis patients. The experimental approaches exploit our experience in molecular genetics, whole genome transcription profiling, promoter target localization, and DNA sequence analysis. The goals are responsive to the RFA since we will analyze P. gingivalis intracellular signaling mechanisms that orchestrate responses to environmental changes. By defining the regulators, the genes they control, and their in vitro and in vivo expression patterns, we can identify conditions that may trigger periodontitis, and target rational therapeutic interventions to the regulatory molecular switches.