The interaction of Chlamydia with the host cytoskeleton

衣原体与宿主细胞骨架的相互作用

• 批准号: 7880695
• 负责人: SCOTT S GRIESHABER
• 金额: $ 36.26万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7880695
• 关键词: Antibodies; Bacteria; Binding; Biochemical; Biological Assay; Blindness; Cell division; Cells; Centrosome; Chimeric Proteins; Chlamydia; Chlamydia Infections; Chlamydia trachomatis; Chromosomal Instability; Chromosomes; Coculture Techniques; Color; Complex; Cytokinesis; Cytoskeleton; DNA; Defect; Density Gradient Centrifugation; Development; Dynein ATPase; Escherichia coli; Expression Library; Failure; Fluorescent Dyes; Genital system; Goals; Growth; Hour; Human; Human Papillomavirus; Human papilloma virus infection; Human papillomavirus 16; Image; Imagery; Immune Sera; Immunoprecipitation; In Vitro; Incidence; Infection; Infertility; Label; Lead; Libraries; Life; Life Cycle Stages; Link; Maintenance; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Mass Spectrum Analysis; Mediating; Membrane; Microscopy; Microtubule Polymerization; Microtubule-Associated Proteins; Microtubule-Organizing Center; Microtubules; Monoclonal Antibodies; Motor; Mus; Oncogenes; Organelles; Organism; Outcome; Pathogenesis; Patients; Pelvic Inflammatory Disease; Play; Process; Protein Binding; Protein Biosynthesis; Protein Fragment; Proteins; Reporting; Research; Role; Scheme; System; Techniques; Tertiary Protein Structure; Time; Trimethoprim-Sulfamethoxazole; United States; Vacuole; Woman; animal tissue; base; cellular imaging; cofactor; cross reactivity; daughter cell; extracellular; genetic regulatory protein; high risk; inhibitor/antagonist; metaplastic cell transformation; migration; mouse development; new therapeutic target; pathogen; protein complex; public health relevance; segregation; trafficking; uptake

DESCRIPTION (provided by applicant): The obligate intracellular bacterium Chlamydia trachomatis is the world's leading cause of preventable blindness and the most common sexually transmitted bacterial pathogen of humans. In the US, the incidence of new cases of chlamydial genital tract infection is approximately 4 million annually, causing severe illness such as pelvic inflammatory disease leading to tubal infertility. Chlamydia replication takes place in a unique membrane bound compartment within the host cell termed the inclusion. One of the earliest steps in inclusion maturation is centripetal migration to the perinuclear region, a process that is conserved among all chlamydial species suggesting an important role in pathogenesis. This migration is chlamydial driven, as chlamydial protein synthesis is required for the recruitment of the microtubule motor protein dynein. Dynein recruitment and activation is required for chlamydial inclusion trafficking to the microtubule organizing center (MTOC) of the cell, which ultimately leads to an intimate association between the inclusion and the host centrosome. The mature chlamydial inclusion remains associated with the centrosomes causing centrosome number defects, spindle defects, and chromosome instability. We hypothesize that the recruitment and activation of dynein is a central mechanism of pathogenesis during chlamydial infection. The focus of this proposal is to identify the chlamydial proteins involved in dynein activation, and investigate the role of dynein and centrosomes in cell to cell spread and cellular transformation. Aim 1 will identify chlamydial proteins that bind to dynein using dynein immunoprecipitation, centrosome purification and microtubule isolation techniques. Centrosomes are dynamic organelles and are actively partitioned into daughter cells during cell division, therefore Aim 2 will investigate the role of dynein and the centrosome in cell-to-cell spread of Chlamydia under persistent and non-persistent growth conditions. Centrosome function is crucial in the maintenance of chromosome fidelity. The unique chlamydial driven interaction between dynein and the inclusion is likely an important factor in the link between chlamydial infection and cervical cancer. Aim 3 will determine the mechanism of chlamydial induced centrosome defects and evaluate these defects in cellular transformation. Additionally, the link between cervical cancer and Chlamydia likely involves an interaction with human papilloma virus (HPV) infection. We will therefore assess the role of the interaction between chlamydial induced centrosome defects and expression of the high risk HPV16 E6 and E7 oncogenes on cellular transformation. PUBLIC HEALTH RELEVANCE: The bacteria Chlamydia infects an estimated 4 million people annually in the United States. Infection with this organism leads to pelvic inflammatory disease, infertility in women and raises the chance of cervical cancer development. Determining the mechanisms of chlamydial intracellular development and its impact on host cell replication machinery will lead to advances in understanding chlamydial disease and offer potential novel therapeutic targets.

描述（由申请人提供）：专性细胞内细菌沙眼性是世界可预防失明的主要原因，也是人类最常见的性传播细菌病原体。在美国，新的衣原体生殖道感染病例的发生率每年约400万，导致严重疾病，例如骨盆炎性疾病，导致输卵管不育。衣原体复制发生在宿主细胞内的独特膜结合室中，称为包容性。纳入成熟的最早步骤之一是中心迁移到核周区，这一过程在所有衣原体物种中都保守，这表明在发病机理中起重要作用。这种迁移是衣原体驱动的，因为募集微管运动蛋白动力蛋白需要衣原体蛋白的合成。 Dynein的募集和激活是经过经纳入纳入的纳入纳入的纳入融合到细胞微管组织中心（MTOC）所必需的，这最终导致包容性与宿主中心体之间建立了亲密的关联。成熟的衣原体包容性仍然与中心体相关，导致中心体数量缺陷，纺锤体缺陷和染色体不稳定性。我们假设动力蛋白的募集和激活是衣原体感染期间发病机理的中心机制。该提案的重点是鉴定与动力蛋白活化有关的衣原体蛋白，并研究动力蛋白和中心体在细胞中对细胞扩散和细胞转化的作用。 AIM 1将使用动力蛋白免疫沉淀，中心纯化和微管分离技术鉴定与动力蛋白结合的衣原体蛋白。中心体是动态细胞器，并在细胞分裂过程中积极分配到子细胞中，因此AIM 2将研究动力蛋白和中心体在持续和非持续生长条件下衣原体在细胞间扩散中的作用。中心体功能对于维持染色体保真度至关重要。动力蛋白与包容性之间独特的衣原体驱动的相互作用可能是衣原体感染与宫颈癌之间联系的重要因素。 AIM 3将确定衣原体诱导的中心体缺陷的机理，并评估这些缺陷在细胞转化中。此外，宫颈癌与衣原体之间的联系可能涉及与人乳头瘤病毒（HPV）感染的相互作用。因此，我们将评估衣原体诱导的中心体缺陷与高风险HPV16 E6和E7 Oncogenes在细胞转化中的相互作用的作用。公共卫生相关性：细菌衣原体每年在美国感染大约400万人。这种生物的感染会导致骨盆炎症性疾病，女性不育症，并增加了宫颈癌发展的机会。确定衣原体细胞内发育的机制及其对宿主细胞复制机制的影响将导致理解衣原体疾病的进步，并提供潜在的新型治疗靶标。