Regulation of bacterial pathogen actin-based motility by host cell kinases

宿主细胞激酶对细菌病原体肌动蛋白运动的调节

• 批准号: 8095076
• 负责人: HERVE F AGAISSE
• 金额: $ 24.83万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8095076
• 关键词: Actins; Affect; Affinity; Biochemical; Biological Assay; CSNK1A1 gene; Cell physiology; Cells; Comparative Study; Complex; Computer-Assisted Image Analysis; Cytoskeleton; Cytosol; Defect; Fluorescence Microscopy; Foundations; Genetic; Human; Integration Host Factors; Knowledge; Lead; Listeria; Mediating; Methodology; Molecular; Mutagenesis; N-terminal; Pathogenesis; Phosphorylation; Phosphotransferases; Play; Preventive Intervention; Process; Production; Protein-Serine-Threonine Kinases; RNA Interference; Regulation; Research Design; Rickettsia; Role; Screening procedure; Seminal; Shigella; Site; Substrate Specificity; Surface; Tail; Therapeutic Intervention; Virulence Factors; Work; base; cell motility; design; innovation; insight; microbial; mimicry; novel; novel strategies; pathogen; research study

DESCRIPTION (provided by applicant): Several intracellular bacterial pathogens, such as Listeria ssp, Rickettsia spp. and Shigella spp., rely on actin-based motility to spread from infected cells to neighboring cells and disseminate within their host. Genetic and biochemical studies have revealed that actin-based motility relies on the production of virulence factors at the bacterial surface that interact with host cell cytoskeleton factors, including the ARP2/3 complex. In spite of these seminal contributions, the host cell factors supporting actin-based motility are still poorly understood. To fill this gap in knowledge, we have developed innovative approaches combining automated fluorescence microscopy and computer-assisted image analysis in order to visualize and quantify bacterial pathogen spread. We have used this newly developed approach to screen the human kinome and identified CK1 and CK2 as host cell kinases required for Listeria spread. We recently demonstrated that, similar to WASP and WAVE2, the affinity of ActA for the ARP2/3 complex is modulated by CK2-mediated phosphorylation, a notion that we refer to as regulatory mimicry. In this proposal we propose genetic and biochemical approaches designed to explore the role(s) of CK1 in the regulation of Listeria actin-based motility (Aim1). In addition, we propose to extend the approach used for investigating Listeria spread to several bacterial pathogens displaying actin-based motility, including Shigella and Rickettsia (Aim2). These exploratory studies may provide important and novel insights both into the mechanisms supporting Listeria, Shigella and Rickettsia pathogenesis and into the fundamental processes involved in the regulation of the actin cytoskeleton. PUBLIC HEALTH RELEVANCE: Various intracellular pathogens have evolved the ability to manipulate host cell processes in order to spread from infected cells into the neighboring cells. Most of he host factors contributing to the cell-to-cell spread of pathogens are unknown. In this proposal, we present our plans to determine and investigate the host cell kinases involved in Listeria spp., Shigella spp. and Rickettia spp. actin-based motility. The proposed approach will contribute to our general understanding of the mechanisms underlying microbial pathogenesis and may therefore constitute the foundation for the rational design of preventive and therapeutic interventions.

描述（由申请人提供）：几种细胞内细菌病原体，例如李斯特氏菌SSP，立克西亚属。和志贺氏菌属于基于肌动蛋白的运动能力从感染细胞传播到邻近细胞，并在其宿主中传播。遗传学和生化研究表明，基于肌动蛋白的运动依赖于与宿主细胞细胞骨架因子相互作用（包括ARP2/3复合物）在细菌表面的产生。尽管有这些开创性的贡献，但支持基于肌动蛋白运动的宿主细胞因素仍然很少了解。为了填补知识的这一空白，我们开发了结合自动荧光显微镜和计算机辅助图像分析的创新方法，以可视化和量化细菌病原体扩散。我们已经使用了这种新开发的方法来筛选人类动物组，并将CK1和CK2鉴定为李斯特菌扩散所需的宿主细胞激酶。我们最近证明，类似于WASP和WAVE2，ACTA对ARP2/3复合物的亲和力是由CK2介导的磷酸化调节的，这一概念是我们称为调节模仿的概念。在此提案中，我们提出了旨在探索CK1在基于李斯特菌肌动蛋白的运动中的作用的遗传和生化方法（AIM1）。此外，我们建议将用于调查李斯特菌扩散的方法扩展到几种细菌病原体，这些病原体显示基于肌动蛋白的运动，包括志贺氏菌和人力素（AIM2）。这些探索性研究可能会为支持李斯特氏菌的发病机理的机制提供重要和新颖的见解，以及与调节肌动蛋白细胞骨架调节有关的基本过程。 公共卫生相关性：各种细胞内病原体已经发展了操纵宿主细胞过程以从感染细胞传播到相邻细胞中的能力。 HE大多数宿主因素导致病原体细胞间传播的因素尚不清楚。在此提案中，我们介绍了我们的计划，以确定和调查与李斯特氏菌Spp。和里克蒂亚属。基于肌动蛋白的运动。所提出的方法将有助于我们对微生物发病机理的机制的一般理解，因此可能构成预防和治疗干预措施的理性设计的基础。