Research Starter Grant: Investigating the biochemical function of Wolbachia pipientis type IV effectors

这个项目的最终目标是确定一种异常广泛的细胞内细菌改变宿主细胞生物学的机制。具体来说，该项目侧重于细胞内细菌沃尔巴克氏体和细菌决定因素的鉴定，操纵真核细胞生物学。该项目利用生物信息学和酵母遗传系统的力量来研究这种不可培养的，遗传上难以处理的细菌。首先，该项目侧重于在生物信息学筛选中鉴定的蛋白质，这些蛋白质可能是沃尔巴克氏体在宿主感染过程中分泌的。这些蛋白质在真核细胞的背景下使用高通量遗传筛选和功能基因组方法进行表征，包括遗传和生化相互作用的鉴定。确定沃尔巴克氏体与其宿主相互作用的机制将有助于在昆虫媒介控制的背景下了解这种共生关系。管状沃尔巴克氏体感染极其广泛的真核宿主，包括蚊病载体伊蚊和按蚊，由于它们与这些昆虫的特殊相关性，最近引起了人们的注意。登革热及其毒性更大的出血热正在成为21世纪严重的公共卫生威胁，预计控制主要病媒蚊子对有效预防流行病至关重要。有趣的是，沃尔巴克氏体感染已被证明可预防伊蚊传播登革热，尽管感染发生的生物学机制尚不清楚。通过表征与真核细胞生物学相互作用的沃尔巴克氏体蛋白，我们可以更好地理解伊蚊-登革热-沃尔巴克氏体相互作用背后的机制，使我们能够更好地预测沃尔巴克氏体感染如何被用来预防蚊子传播疾病。此外，由于沃尔巴克氏体感染如此广泛的昆虫，这项工作的结果将导致对其他感染的比较分析。

The ultimate goal of this project is to identify mechanisms by which an extraordinarily widespread, intracellular bacterium modifies host cell biology. Specifically, this project focuses on the intracellular bacterium Wolbachia and the identification of bacterial determinants that manipulate eukaryotic cell biology. This project leverages the power of bioinformatics and the yeast genetic system to study this unculturable, genetically intractable bacterium. First, the project focuses on proteins, identified in a bioinformatics screen, likely secreted by Wolbachia during the process of host infection. These proteins are characterized in the context of the eukaryotic cell using high-throughput genetic screens and functional genomic approaches including the identification of genetic and biochemical interactions.Identification of the mechanisms by which Wolbachia interacts with its host will contribute to the understanding of this symbiosis in the context of insect vector control. Wolbachia pipientis infects an extremely broad array of eukaryotic hosts, including the mosquito disease vectors Aedes and Anopholes, and have become noteworthy of late due to their specific relevance to these insects. Dengue fever and its more virulent form hemorrhagic fever, are emerging as serious public health threats in the 21st century and controlling the principal vector mosquito is predicted to be critical to the effective prevention of an epidemic. Interestingly, infection by Wolbachia has been shown to prevent transmission of Dengue by Aedes, although the biological mechanisms through which the infection is accomplished are unknown. By characterizing Wolbachia proteins that interact with eukaryotic cell biology, we can better understand the mechanism behind the Aedes-Dengue-Wolbachia interaction, allowing us to better predict how Wolbachia infection may be used to prevent the transmission of diseases by mosquitos. Additionally, because Wolbachia infect such a broad diversity of insects, results from this work will lead to comparative analyses in other infections.