Remodeling of the Host Ubiquitin Landscape by Chlamydia trachomatis

沙眼衣原体重塑宿主泛素景观

基本信息

批准号：
9397352
负责人：
Khavong Pha
金额：
$ 5.67万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-06-01 至 2020-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9397352
关键词：
Affinity Chromatography Antibiotics Apoptosis Bacteria Bacterial Sexually Transmitted Diseases Binding Biochemical Bioinformatics Biological Biology Cell division Cell physiology Cells Cellular biology Centers for Disease Control and Prevention (U.S.)Chlamydia Chlamydia Infections Chlamydia trachomatis Chlamydiales Complex Cytoplasm Data Set Developed Countries Developing Countries Environment Event Eye Infections Eye diseases Future Genetic Goals Grant HIV Health Care Costs Human Human papillomavirus 16 E1 protein Hybrids Infection Infertility Injectable Innate Immune Response Knowledge Learning Life Cycle Stages Ligase Link Listeria monocytogenes Location Mass Spectrum Analysis Membrane Membrane Proteins Modeling Molecular Molecular Biology Mycobacterium tuberculosis Organelles Pathogenesis Post-Translational Protein Processing Prevention Process Protein Export Pathway Proteins Proteome Proteomics Recruitment Activity Respiratory Tract Infections Role Scientist Sexually Transmitted Diseases System Techniques Trachoma Training Type III Secretion System Pathway Ubiquitin Ubiquitin-Activating Enzymes Ubiquitin-Conjugating Enzymes Vaccines Virulence Zinc Fingers bacterial genetics base career disorder prevention experimental study genetic manipulation genital infection human disease insight interest microbial mutant next generation novel pathogen skills therapeutic target tool ubiquitin ligase ubiquitin-protein ligase

项目摘要

PROJECT SUMMARY Chlamydia trachomatis is the leading cause of sexually transmitted diseases, non-congenital infertility, and the blinding eye disease called trachoma. This obligate intracellular bacterium replicates within a membrane bound compartment, the inclusion, where it utilizes a specialized protein export system called the type III secretion system to translocate effector proteins into the cytoplasm of the host cell. A unique subfamily of type III effector proteins the Inclusion membrane proteins (Incs), is inserted into the inclusion membrane, with its N- and C- termini ideally poised at the host-pathogen interface to interact with host proteins and organelles. However, our understanding of the function of Incs has been limited, as they share little homology to each other or to other known proteins, and genetic manipulation of Chlamydia only recently become possible. To define the C. trachomatis Inc-human protein interactome, the Engel lab performed a high-throughput affinity-purification mass spectroscopy screen of 58 predicted Incs. Remarkably, a subset of Incs was identified to target components of the host ubiquitin (Ub) machinery, which prompts the hypothesis that a subset of Chlamydia Incs may modulate host ubiqutylation during infection and that these changes may be critical to the pathogenesis of C. trachomatis infection. Ubiquitylation, which involves the covalent attachment of Ub to a target protein by an E1 Ub-activating enzyme, an E2 Ub-conjugating enzyme, and an E3 Ub ligase, alters the function, stability, or location of the Ub-modified protein within the cell. Modulating protein ubiquitylation during infection is an emerging theme for intracellular pathogens. Indeed, previous studies have established that C. trachomatis infection globally alters the host protein stability; however, how Inc-Ub machinery interactions reshape the host proteome is unknown. The long-term goal of this project is to understand how interactions between specific Incs and the host Ub machinery alters the host Ub landscape and promotes Chlamydia pathogenesis. In aim 1, a newly developed MS-based ubiquitin remnant profiling (URP) approach and bioinformatics will be used to globally define C. trachomatis-induced changes in the host ubiquinome. These changes will be compared to data sets for other intracellular pathogens obtained by a similar pipeline to identify changes in protein ubiquitylation that are specific to Chlamydia infections as well as protein ubiquitylation events that are common targets of intracellular pathogens. In aim 2, a combination of URP, host and bacterial genetics, and cell and molecular biology will be used to identify changes in the host ubiquinome that are dependent on the Inc (CT383) and its interactions with components of host Ub machinery. Together, these studies will provide critical new insights on the mechanisms by which C. trachomatis reprograms the host cell through ubiquitylation to promote pathogenesis. !

项目摘要沙眼衣原体是性传播疾病，非中性不育症和眼科疾病称为沙眼疾病。这种强制性细胞内细菌在膜结合内复制隔室，包含，它利用了称为III型分泌的专业蛋白质导出系统将效应蛋白转移到宿主细胞的细胞质中的系统。 III型效应器的独特亚家族蛋白质包含膜蛋白（INC）插入纳入膜中，其n和c- 末端理想地固定在宿主 - 病原体界面，以与宿主蛋白和细胞器相互作用。但是，我们的了解Incs的功能受到限制，因为它们相互或彼此之间有很少的同源性已知的蛋白质和对衣原体的遗传操纵直到最近才有可能。定义C。气管体INC-HUMAN蛋白质相互作用，Engel Lab进行了高通量亲和力纯化 58个预测INC的质谱屏幕。值得注意的是，INC的子集被确定为目标宿主泛素（UB）机械的组件，这促使假设是衣原体的子集 INC在感染过程中可能调节宿主ubiqutylation，这些变化可能对沙眼梭状芽孢杆菌感染的发病机理。泛素化，涉及UB的共价附件通过E1 UB激活酶，E2 UB偶联酶和E3 UB连接酶的靶蛋白，改变了 UB修饰蛋白在细胞内的功能，稳定性或位置。调节蛋白质泛素化感染是细胞内病原体的新兴主题。确实，以前的研究确定了C. 沙眼感染在全球范围内改变宿主蛋白的稳定性；但是，如何INC-UB机械交互重塑宿主蛋白质组是未知的。该项目的长期目标是了解互动方式在特定的INC和主机UB机械之间，可以改变主机UB景观并促进衣原体发病。在AIM 1中，新开发的基于MS的泛素残留分析（URP）方法和生物信息学将用于全球定义宿主泛素组的沙眼梭菌诱导的变化。这些更改将与通过类似管道获得的其他细胞内病原体的数据集进行比较，以识别特定于衣原体感染和蛋白质泛素化的蛋白质泛素化的变化是细胞内病原体的常见目标。在AIM 2中，URP，宿主和细菌的组合遗传学以及细胞和分子生物学将用于识别宿主泛素体中的变化取决于Inc（CT383）及其与主机UB机械组件的相互作用。在一起，这些研究将提供有关C. c. trachomatis对宿主细胞进行重新编程的机制的关键新见解通过泛素化来促进发病机理。呢