Collaborative Research: How does an intracellular symbiont manipulate host cell biology?

合作研究：细胞内共生体如何操纵宿主细胞生物学？

• 批准号: 1456535
• 负责人: William Sullivan
• 金额: $ 27.1万
• 依托单位: University of California-Santa Cruz
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1456535&HistoricalAwards=false
• 关键词: Collaborative; Research; How; does; intracellular

Wolbachia pipientis is a bacterium that infects a large number of insects, including important human disease vectors such as mosquitos. This bacterium is maternally transmitted and is known to cause a variety of effects in the insect host including manipulation of reproduction and suppression of virus replication. Because Wolbachia is capable of suppressing virus replication in insect hosts, it is being used to decrease the spread of human diseases (so-called "pathogen blocking"). It is critical, therefore, to better understand how Wolbachia infects hosts and alters host cell biology. Although Wolbachia infect upwards of 40% of insects on the planet, there is surprisingly little known about the exact mechanism used by the bacterium to manipulate its host. This is largely due to the fact that Wolbachia occurs inside host cells and cannot be genetically manipulated presently. This project will use a novel approach in this system to identify proteins used by the bacterium to alter host cell biology. By identifying and characterizing these proteins, much will be learned about the basic biology of Wolbachia and the host cell components utilized during infection. Results will also lead to comparative analyses in other symbiotic systems. Undergraduate students from underrepresented groups in science will be trained with project-based research related to this project on Wolbachia.The overarching scientific goal of this project is to identify the mechanisms by which an extraordinarily widespread bacterium interacts with its host. Wolbachia pipientis is an intracellular alpha-proteobacterium that forms symbioses with an extremely broad array of invertebrates, including isopods, nematodes, and insects. Wolbachia is currently of interest with regards to vector control because mosquitoes harboring the bacterium are unable to transmit important human diseases. Because Wolbachia are not currently culturable or genetically tractable, little is known about the molecular basis of interaction with eukaryotic hosts. Wolbachia encodes a type IV secretion system and is thought to secrete proteins (termed "effectors") into the eukaryotic cell to alter the biology of its insect hosts. This project will use a powerful method to identify and study the biochemical function of probable effectors in Wolbachia. The investigators will identify candidate effectors through genomics and bioinformatics. They will use a yeast genetic screen and a secretion assay in a heterologous system to further identify and characterize these candidates. Finally, the researchers will investigate the biochemical function and cellular effects of these proteins in the Drosophila host. These results will be the first comprehensive analysis of Wolbachia secreted substrates and will lead to significant advances in the Wolbachia field and comparative work in other symbiotic strains (such as those infecting filarial nemotodes).

沃尔巴克氏菌是一种感染大量昆虫的细菌，包括重要的人类疾病媒介，如蚊子。这种细菌通过母体传播，已知在昆虫宿主中引起各种影响，包括操纵繁殖和抑制病毒复制。由于沃尔巴克氏菌能够抑制昆虫宿主中的病毒复制，它正被用于减少人类疾病的传播(所谓的“病原体阻断”)。因此，更好地了解沃尔巴克氏菌如何感染宿主并改变宿主细胞生物学是至关重要的。尽管沃尔巴克氏菌感染了地球上40%以上的昆虫，但令人惊讶的是，人们对这种细菌操纵宿主的确切机制知之甚少。这在很大程度上是因为沃尔巴克氏杆菌存在于宿主细胞内，目前还不能通过基因操作。这个项目将在这个系统中使用一种新的方法来识别细菌用来改变宿主细胞生物学的蛋白质。通过鉴定和鉴定这些蛋白质，我们将更多地了解沃尔巴克氏菌的基本生物学和感染过程中所利用的宿主细胞成分。结果还将导致在其他共生系统中进行比较分析。来自科学领域代表性不足群体的本科生将接受与沃尔巴克氏项目相关的基于项目的研究培训。该项目的首要科学目标是确定一种异常广泛的细菌与其宿主相互作用的机制。沃尔巴克氏杆菌是一种细胞内的α-蛋白细菌，它与非常广泛的无脊椎动物形成共生，包括等足类、线虫和昆虫。沃尔巴克氏菌目前在媒介控制方面很受关注，因为携带这种细菌的蚊子不能传播重要的人类疾病。由于沃尔巴克氏菌目前还不是可培养的或遗传上易驯化的，因此人们对与真核宿主相互作用的分子基础知之甚少。沃尔巴克氏菌编码一种IV型分泌系统，被认为能将蛋白质(称为“效应器”)分泌到真核细胞中，以改变其昆虫宿主的生物学。该项目将使用一种强大的方法来鉴定和研究沃尔巴克氏杆菌中可能的效应物的生化功能。研究人员将通过基因组学和生物信息学确定候选效应者。他们将使用酵母遗传筛选和异源系统中的分泌测试来进一步识别和表征这些候选基因。最后，研究人员将研究这些蛋白质在果蝇宿主中的生化功能和细胞效应。这些结果将是对沃尔巴克氏菌分泌底物的首次全面分析，并将导致沃尔巴克氏菌领域的重大进展，以及与其他共生菌株(如感染丝虫线虫的菌株)的比较工作。