Molecular and Cellular Dissection of Campylobacter Jejuni Effector Protein Function

空肠弯曲杆菌效应蛋白功能的分子和细胞解剖

• 批准号: 9298566
• 负责人: Michael E Konkel
• 金额: $ 38.22万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-16 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9298566
• 关键词: ANGPTL2 gene; Abdominal Pain; Actins; Acute; Alpha Cell; Amino Acid Motifs; Antigens; Attenuated; Bacteria; Bacterial Adhesins; Binding; Biological Assay; Blood; Campylobacter; Campylobacter jejuni; Cell Adhesion; Cell physiology; Cell-Matrix Junction; Cells; Cellular Structures; Complex; Coupled; Cytoskeletal Modeling; Cytoskeleton; Cytosol; Data; Development; Devices; Diarrhea; Disease; Disease model; Dissection; Enzyme Inhibitor Drugs; Epidermal Growth Factor Receptor; Epithelial Cells; Event; Fever; Filopodia; Gastrointestinal tract structure; Goals; Guillain-Barré Syndrome; IL8 gene; IQ motif containing GTPase activating protein 1; Immune response; Immunoprecipitation; Infection; Inflammatory Response; Intestines; Knowledge; Laboratories; Leukocytes; Link; Mediating; Membrane; Methods; Microbe; Microfilaments; Modeling; Molecular; Nausea; PTB Domain; Paralysed; Pathway interactions; Phosphorylation; Poliomyelitis; Production; Protein Kinase C; Proteins; Publications; Receptor Activation; Reporting; Research; Research Personnel; Role; Set protein; Signal Pathway; Signal Transduction; Structure; Testing; Therapeutic Intervention; Type III Secretion System Pathway; Virulence; base; cerebral cavernous malformations; chemokine; disease phenotype; enteritis; experimental study; inhibitor/antagonist; insight; mutant; new therapeutic target; novel; pathogen; phosphoproteomics; prevent; protein function; response; uptake

Title: Modulation of Host Cell Function by Campylobacter jejuni Effector Proteins Project Summary Infection with Campylobacter jejuni is responsible for millions of cases of diarrhea per year and is associated with Guillain-Barré syndrome, the leading cause of flaccid paralysis in the post-polio era. Although researchers have surmised for more than two decades that C. jejuni-mediated enteritis is dependent on invasion of the cells lining the gastrointestinal tract and is accompanied by a robust inflammatory response, the pathogen and host responses that contribute to C. jejuni-mediated enteritis are ill-defined. My lab discovered that this pathogen- induced event (cell invasion) requires the delivery of proteins from C. jejuni to the cytosol of host cells. We have designated these secreted proteins the Campylobacter invasion antigens (Cia). The Cia proteins co-opt components of the focal complex (a cell-matrix adhesion structure) and the MEK/ERK signaling pathway to trigger membrane ruffling (lamellipodia and filopodia extensions – cellular protrusions supported by actin filaments). Our overall goal is to identify the mechanism(s) that C. jejuni utilizes to subvert components of the focal complex (FC) to promote cell invasion through cytoskeletal reorganization. Contextually, these studies will provide insight into how microbes modulate cellular signaling pathways to favor pathogen survival and dissemination. We have shown that the CiaC and CiaD effectors are necessary for C. jejuni cellular invasion. Our working hypothesis is that these two effectors contain domains that direct the formation of the Campylobacter invasion complex. More specifically, we hypothesize that CiaC and CiaD co-opt components of the FC and FC- associated proteins that enable the cross-talk (convergence) of cellular pathways and trigger cytoskeletal reorganization and chemokine production (IL-8) in host cells. The precise mechanism(s) that CiaC and CiaD use to modulate cytoskeletal rearrangement and bacterial uptake are not known. Thus, our goal of elucidating how the CiaC and CiaD effectors drive C. jejuni-cell invasion and chemokine induction is essential for understanding the molecular and cellular basis of C. jejuni-mediated enteritis and is a prerequisite for the development of targeted methods to prevent and treat C. jejuni disease. The Specific Aims of this proposal are to: 1) Establish the host cell targets of both CiaC and CiaD; 2) Identify the host cell proteins phosphorylated in a CiaC- and CiaD-dependent manner and determine the role of these phosphorylation events in modulating host cell signaling pathways; and 3) Test whether the C. jejuni CiaC and CiaD effectors are required for disease in piglets and determine if deficiency of either protein results in a unique disease phenotype.

空肠弯曲菌效应蛋白对宿主细胞功能的调节 项目摘要 空肠弯曲杆菌感染每年导致数百万例腹泻，并与 患有格林-巴利综合征，这是后脊髓灰质炎时代导致迟缓性瘫痪的主要原因。尽管研究人员 20多年来一直推测空肠弯曲菌介导的肠炎依赖于细胞的入侵 胃肠道衬里，伴随着强烈的炎症反应，病原体和宿主 导致空肠弯曲菌介导的肠炎的反应是模糊的。我的实验室发现这种病原体- 诱导事件(细胞侵袭)需要将蛋白质从空肠弯曲菌运送到宿主细胞的胞浆中。我们 已将这些分泌蛋白命名为弯曲杆菌入侵抗原(CIA)。中央情报局的蛋白质增选 焦点复合体(一种细胞-基质黏附结构)和MEK/ERK信号通路的组成 触发膜褶皱(片状脂膜和丝状突起--肌动蛋白支持的细胞突起 长丝)。我们的总体目标是确定空肠弯曲菌利用的机制(S)来颠覆 焦点复合体(Fc)通过细胞骨架重组促进细胞侵袭。从背景来看，这些研究 将深入了解微生物如何调节细胞信号通路以有利于病原体的生存和 传播。 我们已经证明，CIAC和CIAD效应是空肠弯曲菌细胞侵袭所必需的。我们的工作 假设这两个效应器包含引导弯曲杆菌入侵形成的结构域。 很复杂。更具体地说，我们假设CIAC和CIAD增选FC和FC的组件- 相关蛋白质，使细胞通路能够串扰(汇聚)并触发细胞骨架 宿主细胞的重组和趋化因子产生(IL-8)。中国工商联和中情局的精确机制(S) 用于调节细胞骨架重排和细菌吸收的作用尚不清楚。因此，我们的目标是澄清 CIAC和CIAD效应如何驱动空肠弯曲菌细胞侵袭和趋化因子诱导是至关重要的 了解空肠弯曲菌介导的肠炎的分子和细胞基础，是研究 开发有针对性的方法来预防和治疗空肠弯曲菌病。 这项建议的具体目的是： 1)建立CIAC和CIAD的宿主细胞靶点； 2)鉴定以依赖ciac和ciad的方式磷酸化的宿主细胞蛋白，并确定 这些磷酸化事件在调节宿主细胞信号通路中的作用；以及 3)检测仔猪是否需要空肠弯曲菌CIAC和CIAD效应器，并确定 任何一种蛋白质的缺乏都会导致一种独特的疾病表型。