POSITIONING OF CAULOBACTER DNA & REGULATORY PROTEINS BY EM TOMOGRAPHY

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 我们已经鉴定了新月柄杆菌染色体的某些区域，其相对于细胞周期动态定位（例如，DNA复制的起点和终点）。 关于这些区域相对于细胞周期时间的定位的高分辨率信息（如何靠近细胞极、中平面以及膜是否相关）是高度感兴趣的。此外，我们已经确定了许多动态本地化的监管和结构蛋白柄杆菌。 我们的本地化研究已经完成了时间推移荧光显微镜下的GFP标记的蛋白质作为一个功能的时间在柄杆菌细胞周期的位置变化。 EM层析成像将提供关于膜定位蛋白质相对于各种细胞表面结构（茎、鞭毛和皮利基部结构和细胞分裂平面）的特定定位的更高分辨率信息。 EM断层扫描的结果将提供关键的洞察力的动态3D定位的调节蛋白在细菌细胞周期的进展和不对称细胞分裂的机制的作用。

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. We have 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, we have identified numerous dynamically localized regulatory and structural proteins in Caulobacter. Our 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.