Ser/Thr protein kinase PknB as target to decrease Streptococcus mutans ecological

描述(申请人提供)：龋齿或龋齿是最常见的传染病之一，影响全世界十分之九的人，在贫困群体中发病率较高。口腔生物膜成员变形链球菌在龋病病因学中的作用已得到很好的证实。因此，管理口腔生物膜中变形链球菌的水平将有利于口腔健康和预防龋病的发生。本项目重点研究了类真核Ser/Thr蛋白激酶PKnB作为抗S。变种人的目标。PKnB是包膜相关应激的重要感受器，并触发相应的细胞反应以增加变形链球菌的应激抗性。最近，我们能够证明一株PnuB缺陷菌株对生态胁迫的敏感性增加。本研究的目的是找出一种最佳的方法来干预依赖PKNB的应激适应。在目标1中，我们建议在生物膜生长的背景下确定pKnB的表达水平和定位。通过这种方法，我们将获得关于变形链球菌生物被膜特异性基因表达的重要信息，并确定PKnB中可获得的结构靶点。在目标2中，我们还将确定下游的PuncB激酶底物，并确定磷酸化级联，以确定变形链球菌特异性PnuB干扰的潜在细胞内靶点。这项研究将扩大对PKnB依赖调控的认识，并有助于开发可能的新的预防龋病的治疗策略。 公共卫生相关性：龋齿是一种常见的传染病，在受影响的人中会造成相当大的不适。变形链球菌在龋病发生中的病因学作用已有很好的文献记载。本项目研究变形链球菌应激反应的一个特定成分，目的是确定干扰应激适应过程的最佳方法。对这一过程的详细了解将为变形链球菌预防龋病的具体干预措施提供重要的新信息。

DESCRIPTION (provided by applicant): Tooth decay or dental caries is one of the most common infectious diseases affecting nine out of ten individuals worldwide with higher rates among underprivileged groups. The role of oral biofilm member Streptococcus mutans in the etiology of dental caries has been well established. Management of S. mutans levels in the oral biofilm would therefore be beneficial for oral health and prevent caries development. This project focuses on the eukaryotic-like Ser/Thr protein kinase PknB as an anti-S. mutans target. PknB is an important sensor of envelope related stress and triggers a respective cellular response to increase S. mutans stress resistance. Recently, we were able to demonstrate the increased susceptibility of a PknB deficient strain towards ecological stress. The goal of the present study is to identify an optimal approach to interfere with PknB dependent stress adaptation. In Aim 1 we propose to determine the levels of pknB expression and localization of PknB in the context of biofilm growth. With this approach we will obtain important information about S. mutans biofilm specific gene expression and identify accessible structural targets in PknB. In Aim 2 we will also identify downstream PknB kinase substrates and determine the phosphorylation cascade to identify potential additional intracellular targets for S. mutans specific PknB interference. This study will expand the knowledge of PknB dependent regulation and help to develop possible novel therapeutic strategies to prevent caries. PUBLIC HEALTH RELEVANCE: Dental caries is a common infectious disease causing considerable discomfort in affected individuals. The etiological role of Streptococcus mutans in caries development has been well documented. This project investigates a specific component of S. mutans stress response with the goal of determining the optimal approach to interfere with the stress adaptation process. A detailed understanding of this process will provide important new information for S. mutans specific interventions to prevent dental caries.