Functional characterization of early Chlamydia effectors

早期衣原体效应器的功能特征

• 批准号: 10406259
• 负责人: Raphael H Valdivia
• 金额: $ 46.42万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-01 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10406259
• 关键词: Animal Model; Bacteria; Binding; Cell Culture Techniques; Cells; Chlamydia; Chlamydia Infections; Chlamydia trachomatis; Chronic; Epithelial; F-Actin; Genetic; Goals; Immune signaling; Impairment; Infection; Infertility; Inflammation; Innate Immune Response; Intercellular Junctions; Interferon Type I; Lead; Mammalian Cell; Mediating; Membrane; Modification; Molecular; Molecular Genetics; Morbidity - disease rate; Mutation; Pathogenicity; Pathology; Pelvic Inflammatory Disease; Phosphatidylinositols; Phosphotransferases; Play; Process; Proteins; Proteomics; Regulation; Reproductive Health; Research; Role; Sexual Transmission; Signal Pathway; Signal Transduction; Surface; System; Testing; Therapeutic Intervention; Tissues; Vacuole; Virulence Factors; Woman; Work; functional group; in vivo; mutant; novel; pathogen; public health relevance; response; targeted treatment; tool; urogenital tract

Project Description/Summary/Abstract The obligate intracellular bacterium Chlamydia trachomatis is a widely disseminated obligate pathogen that infects epithelial surfaces of the urogenital tract, leading to severe sequela such as pelvic inflammatory disease, and infertility. During the initial stages of infection Chlamydia likely delivers between 10-15 separate Type III secreted (T3S) “effector” proteins into the target host cell. We hypothesize that these Chlamydia effector proteins work co-operatively to temporally reprogram signaling pathways and cytoskeletal functions to promote cell invasion and establish a nascent pathogenic vacuole (“inclusion”) In our first aim, we propose to apply emerging genetic and molecular genetic tools in Chlamydia to define the role of early effectors play in invasion, inclusion maturation and in vivo infections. We also propose to perform an epistatic analysis of combinations of effector mutants to group effector by functional groups and to prioritize a detailed molecular characterization of effector that are central signaling nodes. In our second aim, we focus on the characterization of Tepp, an effector that plays important roles in reprograming signaling pathways, including those involved in innate immune responses. We propose to define the mechanism of activation of Class I phosphoinositide 3 kinases (PI3K) at nascent inclusions and the role these activities play in the regulation of membrane dynamics and activation of Type I interferon responses. In parallel, we will identify the molecular players that lead to the Tepp- mediated activation and modification of Eps8, a regulator of Rac1 activity and cell-cell junctions, by applying “proximity proteomics” approaches. Overall, our research plan seeks to perform both a systems levels assessment of the function of early effectors and a detailed molecular characterization of mechanism of action for selected effectors. The overall goal is to define the molecular basis of how specific effector(s) function, identify how signaling pathways are re-programmed and how they all ultimately contribute to Chlamydia survival in host tissues and the induction of pathology.

项目描述/摘要/摘要 专性细胞内细菌沙眼衣原体是一种广泛传播的专性 一种感染泌尿生殖道上皮表面的病原体，导致严重的后遗症，如盆腔炎 炎症性疾病和不孕症。在感染的最初阶段，衣原体可能会 10-15个单独的III型分泌（T3 S）“效应”蛋白进入靶宿主细胞。我们 假设这些衣原体效应蛋白协同工作以暂时重编程 信号通路和细胞骨架功能，以促进细胞侵袭和建立新生的 致病液泡（“内含物”） 在我们的第一个目标中，我们建议在衣原体中应用新兴的遗传和分子遗传工具， 明确早期效应子在侵袭、包涵体成熟和体内感染中的作用。我们也 建议通过以下方式对效应突变体的组合进行上位性分析，以将效应子分组： 功能组，并优先考虑效应子的详细分子表征， 信令节点。 在我们的第二个目标中，我们专注于表征Tepp，一个效应器，发挥重要作用， 在重新编程信号通路，包括那些参与先天免疫反应。我们提出 为了确定I类磷酸肌醇3激酶（PI 3 K）在新生时的活化机制， 包涵体和这些活动在调节膜动力学和激活 I型干扰素反应。与此同时，我们将确定导致特普的分子参与者- 介导的激活和修饰Eps 8，一种Rac 1活性和细胞-细胞连接的调节因子， 应用“邻近蛋白质组学”方法。 总的来说，我们的研究计划旨在对 早期效应物和所选效应物的作用机制的详细分子表征。 总体目标是定义特异性效应子如何发挥功能的分子基础， 信号通路被重新编程，以及它们如何最终有助于衣原体在 宿主组织和病理诱导。

项目成果

A renewed tool kit to explore Chlamydia pathogenesis: from molecular genetics to new infection models.

• DOI: 10.12688/f1000research.18832.1
• 发表时间: 2019-01-01
• 期刊: F1000Research
• 作者: Dolat, Lee;Valdivia, Raphael H
• 通讯作者: Valdivia, Raphael H

Chlamydia repurposes the actin-binding protein EPS8 to disassemble epithelial tight junctions and promote infection.

• DOI: 10.1016/j.chom.2022.10.013
• 发表时间: 2022-12-14
• 期刊: CELL HOST & MICROBE
• 影响因子: 30.3
• 作者: Dolat, Lee;Carpenter, Victoria K.;Chen, Yi-Shan;Suzuki, Michitaka;Smith, Erin P.;Kuddar, Ozge;Valdivia, Raphael H.
• 通讯作者: Valdivia, Raphael H.

The acetylase activity of Cdu1 regulates bacterial exit from infected cells by protecting Chlamydia effectors from degradation.

Cdu1 的乙酰化酶活性通过保护衣原体效应子免遭降解来调节细菌从受感染细胞中排出。

• DOI: 10.1101/2023.02.28.530337
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Bastidas,RobertJ;Kędzior,Mateusz;Davidson,RobertK;Walsh,StephenC;Dolat,Lee;Sixt,BarbaraS;Pruneda,JonathanN;Coers,Jörn;Valdivia,RaphaelH
• 通讯作者: Valdivia,RaphaelH