POSITIONING OF CAULOBACTER DNA & REGULATORY PROTEINS BY EM TOMOGRAPHY

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。麦克亚当斯实验室已经确定了新月弯杆菌染色体的某些区域，这些区域相对于细胞周期动态定位(例如，DNA复制的起点和终点)。关于这些区域的位置与细胞周期时间(细胞极点、中面和膜是否相关)之间的高分辨率信息具有很高的兴趣。此外，他们还在Caulbacter中鉴定了许多动态定位的调节和结构蛋白。它们的定位研究是通过时间推移荧光显微镜完成的，根据GFP标记的蛋白质在Caulbacter细胞周期中的位置随时间的变化而变化。EM断层扫描将提供关于膜定位蛋白相对于各种细胞表面结构(柄、鞭毛和毛基结构以及细胞分裂平面)的特定位置的更高分辨率信息。EM断层扫描结果将为调控蛋白的动态3D定位在细菌细胞周期进展中的作用以及细胞不对称分裂的机制提供关键的见解。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The McAdams lab has identified certain regions of the Caulobacter crescentus chromosome that are dynamically localized versus the cell cycle (e.g., the origin and terminus of DNA replication). High resolution information on positioning of these regions versus cell cycle time(how near the cell poles, the mid-plane, and whether membrane associated) is of high interest. In addition, they have identified numerous dynamically localized regulatory and structural proteins in Caulobacter. Their localization studies have been done by time-lapse fluorescent microscopy following the changing position of GFP-tagged proteins as a function of time in Caulobacter cell cycle. EM tomography will provide higher resolution information on specific positioning of membrane-localized proteins versus various cell surface structures (stalk, flagella and pili base structures and cell division plane). The EM tomography results will provide key insights into the role of dynamic 3D positioning of regulatory proteins in progression of the bacterial cell cycle and into the mechanism of asymmetric cell division.