Type III Secretion Translocon Structure and Function

III型分泌易位子的结构和功能

• 批准号: 8701601
• 负责人: Arne Rietsch
• 金额: $ 23.78万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8701601
• 关键词: Aeromonas hydrophila; Antibiotic Resistance; Bacteremia; Bacteria; Biological Assay; Burn injury; Bypass; Cell membrane; Cells; Corneal Injury; Cysteine; Cytoplasm; Data; Development; Disease; Docking; Drug Targeting; Engineering; Erythrocyte Membrane; Genetic; Genetic Techniques; Heart; Hospitals; Immune system; In Vitro; Infection; Injection of therapeutic agent; Maps; Membrane; Molecular; Mutation; Nature; Needles; Nosocomial Infections; Pasteurella pseudotuberculosis; Patients; Pneumonia; Process; Protein-Protein Interaction Map; Proteins; Pseudomonas aeruginosa; Resistance; Severity of illness; Structure; Surface; Syringes; Techniques; Testing; Therapeutic Intervention; Type III Secretion System Pathway; Ventilator; Virulence Factors; Work; antimicrobial; crosslink; design; disulfide bond; drug development; efflux pump; hybrid protein; in vivo; interest; loss of function; mortality; mutant; neutrophil; pathogen; prevent; public health relevance; research study; resistance mechanism; success

DESCRIPTION (provided by applicant): P. aeruginosa is a common cause of hospital-acquired infections. It uses a type III secretion system to directly inject effector proteins into targeted host cells, in particular neutrophils, thereby preventing clearance by the host immune system. Injection of effector proteins involves a specialized structure called the translocon, which consists of a pore in the host cell membrane that interacts with the tip of the type III secretion needle. While the translocon is critical for type III secretion function, it has been difficult to study biochemically. Here we propose to use a genetic technique to identify critical translocator interactions and categorize them according to their function in the translocation process (translocator insertion, pore formation, docking of the tip to the pore and triggering of effector secretion). Identifying translocator contacts that are critical for function not only increases our understanding of the translocon, it also identifies potential targets for therapeutic intervention that lie outside of the bacterial cell and are therefore not shielded by conventional antibiotic resistance mechanisms, such as the outer membrane or efflux pumps.

描述(申请人提供)：铜绿假单胞菌是医院获得性感染的常见原因。它使用III型分泌系统将效应蛋白直接注入目标宿主细胞，特别是中性粒细胞，从而防止宿主免疫系统清除。效应蛋白的注射涉及一种称为易位子的特殊结构，它由宿主细胞膜上的一个孔组成，该孔与III型分泌针的尖端相互作用。虽然转运子对III型分泌功能至关重要，但生物化学研究一直很困难。在这里，我们建议使用基因技术来识别关键的转运子相互作用，并根据它们在转位过程中的功能(转运子插入、孔形成、顶端对接到孔和触发效应器分泌)进行分类。识别对功能至关重要的转位接触不仅增加了我们对转位的了解，还识别了潜在的治疗靶点 位于细菌细胞外的干预，因此不能被传统的抗生素耐药机制，如外膜或外排泵所屏蔽。