RUI: The Role of Outer Membrane Agglutinins In Bacterial Colonization

智慧价值某些细菌可以在真核生物的外部、粘膜和细胞内定植。细菌外表面的蛋白质促进了细菌的定植，这些蛋白质介导了与真核细胞、固体表面或其他细菌的相互作用。这个项目的重点是凝集素，这是一个外膜蛋白家族，由广泛的革兰氏阴性细菌表达，从人类肠道定殖者到昆虫的细胞内定殖者。本实验室已证实，耐热凝集素1(Hra1)是具有优异定植能力的大肠埃希菌的辅助定植因子。该项目将使用培养的上皮细胞和线虫秀丽线虫作为模型系统，研究该蛋白质与其他表面因素(如菌毛)的相互作用。这项研究还将识别和验证Hra1和其他肠道定殖者的相关蛋白中与定植相关的基序。研究将扩展到沃尔巴克氏菌表面蛋白(WSP)。WSP是一种凝集素，被认为有助于昆虫和线虫细胞内细菌的定位和细菌间的相互作用。为了克服培养和操作沃尔巴克氏菌的困难，我们建议通过在大肠杆菌中表达WSP来研究WSP，并应用我们正在使用的检测大肠杆菌表面蛋白的方法来阐明WSP的功能。这项工作将促进对细菌之间以及细菌与其真核宿主之间相互作用的理解。广泛影响：从该项目中获得的知识将增加对允许细菌在其他生物体上或体内生存的特定分子相互作用的理解。这项研究将在哈弗福德学院进行，这是一家小型文科机构，主要由有指导的本科生研究人员进行。PI教授微生物学和遗传学入门，激发了许多本科生对细菌学的兴趣，每年将为3-8名学生提供与该项目相关的研究机会，其中包括女性和其他在科学界代表性不足的群体。此外，每年将有大约30名学生在以探究为基础的实验室课堂上接受教学，以检验与该项目相关的一些假设。与过去一样，一些接受过这样培训的学生将与PI一起参加为美国K-12学生或西非学生和教师举办的微生物学研讨会。

Intellectual MeritCertain bacteria can colonize external, mucosal and intracellular sites of eukaryotes. Colonization is promoted by proteins on the outer surface of bacteria that mediate interactions with eukaryotic cells, solid surfaces, or other bacteria. This project focuses on the agglutinins, a family of outer membrane proteins expressed by a broad repertoire of Gram-negative bacteria ranging from human intestinal colonizers to intracellular colonizers of insects. Our laboratory has shown that the Heat Resistant Agglutinin 1 (Hra1) is an accessory colonization factor in Escherichia coli strains with exceptional colonization abilities. This project will examine interplay of this protein with other surface factors, such as fimbriae, using cultured epithelial cells and the nematode worm Caenorhabditis elegans as model systems. The research will additionally identify and validate colonization-associated motifs within Hra1 and related proteins from other intestinal colonizers. The investigations will be extended to Wolbachia Surface Proteins (Wsps). Wsps are agglutinins hypothesized to contribute to localization and interbacterial interactions of bacteria that reside inside the cells of insects and nematodes. To get around difficulties with culturing and manipulating Wolbachia, we propose to study the Wsps by expressing them in E. coli and to apply the methods we are using to examine E. coli surface proteins towards elucidating Wsp function. This work will advance the understanding of interactions among bacteria as well between bacteria and their eukaryotic hosts.Broader impact: Knowledge gained from this project will increase understanding of specific molecular interactions that allow bacteria to live on or in other organisms. The research will be conducted at Haverford College, a small liberal arts institution and will largely be performed by mentored undergraduate researchers. The PI teaches microbiology and introductory genetics, stimulating interest in bacteriology in a number of undergraduates and will offer 3-8 students each year, including women and other groups underrepresented in the sciences, research opportunities associated with the project. Additionally, approximately 30 students a year will be taught in inquiry-based lab classes that test some hypotheses associated with the project. As in the past, some students so trained will work with the PI in microbiology workshops for K-12 students in the US or for students and Faculty in West Africa.