Vector and Host Contributions to the Regulation of E. chaffeensis Gene Expression

载体和宿主对查菲埃里希体基因表达调节的贡献

• 批准号: 8059879
• 负责人: ROMAN R. GANTA
• 金额: $ 6.19万
• 依托单位: KANSAS STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-12-15 至 2012-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8059879
• 关键词: Address; Anaplasma; Anaplasma phagocytophilum; Anaplasmataceae; Antigens; Arthropods; Bacteria; Biological; Bovine Anaplasmosis; Case Study; Cells; Chlamydia trachomatis; Complex; Coupled; Disease; Ehrlichia; Ehrlichia chaffeensis; Ehrlichiosis; Environment; Family; Gene Expression; Gene Expression Regulation; Genes; Genetic Materials; Genetic Transcription; Genome; Goals; Growth; Health; Host Defense; Human; Immune response; In Vitro; Infection; Infection Control; Knock-out; Knowledge; Life; Life Cycle Stages; Maps; Membrane Proteins; Methods; Molecular; Morphology; Mus; Mutation; Organism; Outcome Study; Pathogenesis; Pattern; Process; Protein Biosynthesis; Proteins; Protocols documentation; Regulation; Reporting; Rickettsia; Rickettsiales; Signal Transduction; System; Tick Control; Ticks; Tissue-Specific Gene Expression; Translations; Vertebrates; abstracting; base; gene function; human granulocytic ehrlichiosis; invertebrate host; knockout gene; macrophage; meetings; monocyte; mortality; novel; pathogen; pathogenic bacteria; promoter; protein E; protein expression; research study; response; tool; vector

PROJECT ABSTRACT: Illnesses caused by the tick-borne rickettsiales of the genera Ehrlichia and Anaplasma have been a growing health concern in recent years. These illnesses include human monocytic ehrlichiosis, human granulocytic ehrlichiosis and human granulocytic anaplasmosis which are caused by Ehrlichia chaffeensis, E. ewingii and Anaplasma phagocytophilum, respectively. Despite the complex cellular environment of arthropods and vertebrates having sophisticated systems of defense, the rickettsiales evolved strategies to evade host clearance. For example, E. chaffeensis in macrophages and tick cells differ significantly in their morphology with distinct patterns of protein expression. The expression patterns include the differential expression from the p28-Omp multigene locus which contains 22 genes. The p28 Omp gene expression is restricted primarily to the p28-Omp gene 19 in macrophages, whereas in tick cells the expressed antigen is predominantly from the p28-Omp gene 14. Our studies also demonstrate that the differential expression in vertebrate and tick cells contributes to the altered host response, such as the delayed clearance in a vertebrate host for tick cellderived E. chaffeensis compared to that for macrophage-derived bacteria. This study will focus on the basic understanding of the regulation of gene expression from the p28-Omp multigene locus. The central hypothesis of this study is that the transcription from the p28-Omp locus in E. chaffeensis is differentially regulated in response to environmental signals and that the host cell-specific gene expression is essential for pathogen¿s survival in vertebrate and tick cell environments. Specific aims of this study are; 1) establish in vitro transcription and translation system for mapping Ehrlichia promoters, 2) map transcriptional machinery in differentially expressed genes 14 and 19 of the p28-Omp locus, 3) evaluate the contributions of macrophage and tick cell environments for differential expression from genes 14 and 19, and 4) evaluate knockout mutations for the p28-Omp genes 14 and 19 to assess the biological relevance of host cell specific expression. The results from this study will provide important information for understanding E. chaffeensis gene regulation and how the rickettsiale in macrophages and tick cells respond to the loss of expression from a differentially expressed protein.

项目摘要： 由ehrlichia和Anaplasma属的tick传播的立克菌引起的疾病一直在不断增长 近年来健康关注。这些疾病包括人类的单核细胞性卵性，人粒细胞 eHrllichios和人类粒细胞肿瘤病是由Ehrlichia Chaffeensis，E.Ewingii和Ewingii和E. ewingii和 分别是吞噬吞噬的。尽管节肢动物和 脊椎动物具有复杂的防御系统，立克人士发展了逃避主机的策略 清除。例如，巨噬细胞和壁虱细胞中的小杂志在其形态上有明显的不同 具有蛋白质表达的不同模式。表达模式包括与 p28-pomp的多基因基因座，其中包含22个基因。 P28 OMP基因表达受到限制 到巨噬细胞中的p28-om基因19，而在壁虱细胞中，表达的抗原主要来自 P28-OM基因14。我们的研究还表明，脊椎动物和tick中的差异表达 细胞有助于改变的主机响应，例如脊椎动物的脊椎动物中的延迟间隙 与巨噬细胞衍生的细菌相比，小杂菌菌。这项研究将集中于基本 了解来自p28-om多基因基因座基因表达的调节。中心假设 这项研究的是，Chaffeensis中P28-OM基因座的转录受到差异调节 对环境信号的反应，并且宿主细胞特异性基因表达对于病原体至关重要 在脊椎动物和壁虱环境中的生存。这项研究的具体目的是； 1）在体外建立 映射Ehrlichia启动子的转录和翻译系统，2）地图转录机械 P28-OM基因座的不同表达基因14和19，3）评估巨噬细胞的贡献 以及与基因14和19的差异表达的滴答细胞环境，以及4）评估基因敲除 P28-OM基因14和19的突变，以评估宿主细胞特异性表达的生物学相关性。 这项研究的结果将提供重要的信息，以理解Chaffeensis基因调节 以及巨噬细胞和壁虱细胞中的立克索尔如何应对差异表达的丧失 表达的蛋白质。