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

该项目的最终目标是确定一种异常广泛的细胞内细菌修改宿主细胞生物学的机制。具体地说，这个项目的重点是细胞内的沃尔巴克氏菌和识别操纵真核细胞生物学的细菌决定因素。这个项目利用生物信息学和酵母遗传系统的力量来研究这种无法培养、遗传上难以克服的细菌。首先，该项目将重点放在生物信息学筛查中确定的蛋白质上，这些蛋白质可能是由沃尔巴克氏菌在宿主感染过程中分泌的。这些蛋白质在真核细胞中使用高通量遗传筛选和功能基因组学方法进行表征，包括鉴定遗传和生化相互作用。确定沃尔巴克氏菌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理解这种共生。细管沃尔巴克氏菌感染的真核宿主非常广泛，包括蚊子疾病的媒介伊蚊和Anopholes，并因其与这些昆虫的特殊相关性而最近引起人们的注意。登革热及其更致命的形式出血热正在成为21世纪的严重公共卫生威胁，控制主要媒介蚊子被预测为有效预防疫情的关键。有趣的是，沃尔巴克氏菌感染已被证明可以防止登革热通过伊蚊传播，尽管感染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菌蛋白，我们可以更好地了解伊蚊-登革热-沃尔巴克氏菌相互作用背后的机制，使我们能够更好地预测沃尔巴克氏菌感染可能如何被用来防止蚊子传播疾病。此外，由于沃尔巴克氏菌感染的昆虫种类如此广泛，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.