Dissecting the molecular players essential for polar growth in Agrobacterium

这个项目有可能改变我们对细菌生长的理解。在细菌生长过程中，细胞壁通常被理解为通过分散生成新的壁材料而扩大。然而，现在人们知道，一些重要的农业和病原菌在生长过程中通过在细胞的末端(极点)添加新材料来扩大细胞壁。这个项目将阐明农杆菌的极地生长机制，以及极地生长与细胞分裂相结合的方式。该项目将提供植物科学方面的博士后研究员和研究生高级培训，PI将通过在博物馆和大学Calday公共开放参观活动期间的讲座进行公共宣传。国际和平研究所还将向非理科专业的本科生教授一门课程，内容是《植物的秘密生活》，重点是社会、环境和经济方面。该项目将阐明细菌的极性生长机制，解决以下具体问题：1)定义组成农杆菌极性生长机制的蛋白质，在酵母双杂交筛选中使用全长和农杆菌特有的FTSA、FtsZ、POPZ、PopJ、LDT-0845和TipN亚域作为诱饵，以确定新的相互作用伙伴。2)利用核糖开关技术对FTSA、FtsZ、Popz、PodJ、LDT-0845和TipN的细胞进行急性耗竭，并使用延时荧光去卷积和高分辨率结构照明显微镜(SIM)来确定其主要表型。3)农杆菌TipN-at作为潜在的极地生长组织中心的作用。

This project has the potential to change our understanding of growth in bacteria. During bacterial growth the cell wall is typically understood to enlarge by the dispersed generation of new wall material. However, it is now known that some important agricultural and pathogenic bacteria expand their cell walls during growth by adding new material at the ends (poles) of the cells. This project will elucidate the mechanisms of polar growth in Agrobacterium and the manner in which polar growth is integrated with cell division. The project will provide a postdoctoral fellow and graduate student advanced training in plant science, and the PI will engage in public outreach with lectures at museums and during the University's CalDay public open house event. The PI will also teach a course a course on "The Secret Life of Plants" to non-science major undergraduates that emphasizes social, environmental and economic aspects. The project will elucidate the mechanisms of polar growth in bacteria, addressing the following specific issues: 1) Defining the proteins that comprise the Agrobacterium polar growth machinery, using full-length and Agrobacterium specific sub-domains of FtsA, FtsZ, PopZ, PopJ, LDT-0845, and TipN as bait in yeast two hybrid screens to identify novel interacting partners. 2) Genetic depletion of essential Agrobacterium cell cycle factors to determine their primary phenotypes, using riboswitch technology to acutely deplete cells of FtsA, FtsZ, PopZ, PodJ, LDT-0845, and TipN and using time-lapse fluorescence deconvolution and high resolution structured illumination microscopy (SIM). 3) The role of Agrobacterium TipN-At as a potential polar growth organizing center.