The molecular basis of bacteria-host interaction Drosophila melanogaster

细菌与宿主相互作用的分子基础 果蝇

• 批准号: 7220804
• 负责人: LAURA R SERBUS
• 金额: $ 4.68万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA CRUZ
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-01-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7220804
• 关键词: Actins; Animal Model; Arthropods; Bacteria; Bacterial Infections; Binding; Biological Assay; Blindness; Candidate Disease Gene; Cells; Centrifugation; Communities; Cytoskeleton; Data; Disease; Disruption; Drosophila genus; Drosophila melanogaster; Embryo; Endoplasmic Reticulum; Ensure; Genes; Genetic; Goals; Human; Image; Intracellular Transport; Life; Localized; Mass Spectrum Analysis; Membrane; Methods; Microbe; Microtubule Proteins; Microtubules; Modification; Molecular; Motor; Nematoda; Onchocerca volvulus; Oocytes; Organelles; Population; Process; Proteins; Public Health; Recruitment Activity; Research; Sedimentation process; Testing; Time; Tissues; Vacuole; Wolbachia; Work; base; cell motility; egg; genetic regulatory protein; human disease; insight; interest; mutant; reproductive

DESCRIPTION (provided by applicant): One of the most successful intracellular bacteria is Wolbachia, harbored by thousands of arthropods and nematodes. Wolbachia have recently been shown cause blindness in humans upon release from their nematode host Onchocerca volvulus. Wolbachia have also been shown to infect the well-established model organism Drosophila melanogaster. Wolbachia rely upon maternal transmittance to spread throughout nematode and arthropod populations. To ensure maternal inheritance, Wolbachia must become incorporated into the reproductive germline cells. This study will be the first to define the molecular basis of Wolbachia inheritance in Drosophila. Three specific aims will be pursued: Aim 1 will define the mechanism used by Wolbachia to promote maternal inheritance. Preliminary work indicates that Wolbachia co-localize at the oocyte posterior with germline determinants that give rise to germline cell formation in the embryo. We hypothesize that Wolbachia co-opt the two-step process used for localization of germline determinant proteins: microtubule- based transport and actin-based anchoring at the posterior cortex. To define the mechanism of Wolbachia posterior localization, we will test Wolbachia distribution in mutant backgrounds that disrupt germline determinant localization. Aim 2 will probe molecular interactions between Wolbachia and host. We hypothesize that Wolbachia modify the host protein composition of the vacuole membrane surrounding them to promote maternal inheritance of Wolbachia. To identify Wolbachia-induced modifications to the vacuole membrane, we will perform gradient sedimentation of organelles followed by mass spectrometry. Aim 3 will test the impact of Wolbachia-induced vacuole modifications on Wolbachia inheritance. Candidate host genes likely to elucidate linkage between Wolbachia and posterior determinants or the host cytoskeleton will be the top priority. Wolbachia localization, motility and quantity will be examined in oocytes mutant for those candidate genes. A subset of functionally host relevant proteins will be tested with deletion constructs and GFP-fusions to identify the domains important for Wolbachia localization as well as when/where the host proteins are recruited to Wolbachia-containing vacuoles. This is an exciting time to enter the Wolbachia research. With the direct relevance to human disease and the potential to explore host-microbe interactions from the perspective of Drosophila, studies of Wolbachia are of increasing interest to public health. The long-term goal of this project is to contribute to the community-wide objective of finding new, more effective strategies for curing bacterial diseases.

描述（由申请人提供）：最成功的细胞内细菌之一是沃尔巴克氏体，由数千种节肢动物和线虫寄生。沃尔巴克氏体最近被证明在从其线虫宿主旋绕盘尾丝虫释放后会导致人类失明。沃尔巴克氏体也被证明可以感染成熟的模式生物黑腹果蝇。沃尔巴克氏体依靠母体传播在线虫和节肢动物种群中传播。为了确保母体遗传，沃尔巴克氏体必须被纳入生殖生殖细胞。这项研究将是第一个定义果蝇中沃尔巴克氏体遗传的分子基础。将追求三个具体目标：目标1将定义沃尔巴克氏体用于促进母性遗传的机制。初步工作表明，沃尔巴克氏体共定位在卵母细胞后部与生殖系决定因素，引起生殖系细胞形成的胚胎。我们假设，沃尔巴克氏体增选用于生殖系决定簇蛋白定位的两步过程：微管为基础的运输和肌动蛋白为基础的锚定在后皮质。为了确定沃尔巴克氏体后定位的机制，我们将测试沃尔巴克氏体分布在突变体背景下，破坏生殖系决定簇定位。目的二是研究沃尔巴克氏体与宿主的分子相互作用。我们推测，沃尔巴克氏体修改其周围的液泡膜的宿主蛋白质组成，以促进沃尔巴克氏体的母系遗传。为了鉴定沃尔巴克氏体诱导的液泡膜修饰，我们将进行细胞器的梯度沉降，然后进行质谱分析。目的3将测试沃尔巴克氏体诱导的空泡修饰对沃尔巴克氏体遗传的影响。候选宿主基因可能阐明沃尔巴克氏体和后决定簇或宿主细胞骨架之间的联系将是重中之重。在这些候选基因突变的卵母细胞中检测沃尔巴克氏体的定位、运动性和数量。将用缺失构建体和GFP融合物测试功能宿主相关蛋白质的子集，以鉴定对沃尔巴克氏体定位重要的结构域以及宿主蛋白质何时/何处被募集到含沃尔巴克氏体的空泡。这是进入沃尔巴克氏体研究的激动人心的时刻。由于与人类疾病的直接相关性以及从果蝇的角度探索宿主-微生物相互作用的潜力，Wolbachia的研究对公共卫生越来越感兴趣。该项目的长期目标是为整个社区的目标做出贡献，即找到治疗细菌性疾病的新的、更有效的策略。