ELECTRON TOMOGRAPHIC CHARACTERIZATION OF CIS-GOLGI CISTERNAL ASSEMBLY

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。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。所列机构为中心，不一定是研究者所在机构。Scherffelia dubia的高尔基体已成为研究池成熟的高尔基体内运输的模型。这条蛇在它的高尔基体中产生鳞片。鳞片只在高尔基池内可见，而在周围的小泡中看不到。这一观察结果是脑池成熟假说的基石。为了进一步测试这一假设的结构预测，我们已经产生了3-D高尔基重建高压冷冻和冷冻替代细胞在约6 nm的分辨率，使用电子断层扫描。这种分析允许详细的表征和量化的结构参与顺式脑池组装。每个过渡ER域平均显示9个约70 nm的COPII型出芽曲线。大约30%的COPII型囊泡在过渡性ER和最顺式的高尔基池之间的空间中形成3-5个囊泡的栓系簇。在附着到C2脑池后，栓系的COPII型囊泡似乎融合以产生脑池起始物，该脑池起始物通过与单独的COPII型囊泡融合而生长成新的脑池。假定的对接/融合复合物使囊泡在与脑池膜融合之前变形成锥形。两种类型的接头复合物连接生长的顺式池，一种类型连接中间池和反式池。披衣细胞和高等植物的高尔基体表现出相同的功能，这表明他们也交通池成熟。 Donohoe，BS，ZR German，M. Gerl和LA Staehelin（2005）在Scherffelia dubia，Chlamyelae reinhardtii和高等植物中顺式高尔基体池状装配和相关COPII囊泡出芽和融合事件的电子断层扫描表征。综述Ce的分子生物学

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 Golgi of Scherffelia dubia has become a model for the study of intra-Golgi transport by cisternal maturation. This alga produces scales in its Golgi stacks. Scales are seen only inside the Golgi cisternae and not in the surrounding vesicles. This observation is a cornerstone of the cisternal maturation hypothesis. To further test the structural predictions of this hypothesis, we have produced 3-D Golgi reconstructions of high-pressure frozen and freeze-substituted cells at approximately 6 nm resolution using electron tomography. This analysis has allowed a detailed characterization and quantification of the structures involved in cis-cisterna assembly. Each transitional ER domain displays an average of nine approximately 70 nm COPII-type budding profiles. Approximately 30% of the COPII-type vesicles form tethered clusters of 3-5 vesicles in the space between the transitional ER and the cis-most Golgi cisterna. After attachment to the C2 cisterna, the tethered COPII-type vesicles appear to fuse to create cisternal initials that grow into a new cisterna by fusion with individual COPII-type vesicles. Putative docking/fusion complexes deform vesicles into a cone shape before they fuse with cisternal membranes. Two types of linker complexes connect growing cis-cisternae and one type connects medial- and trans-cisternae. Chlamydomonas and higher plant Golgi exhibit the same features, suggesting that they too traffic by cisternal maturation. Donohoe, BS, ZR Gergely, M. Gerl and LA Staehelin (2005) Electron tomographic characterization of cis-Golgi cisternal assembly and associated COPII vesicle budding and fusion events in Scherffelia dubia, Chlamydomonas reinhardtii, and higher plants. In review Mol.Biol. of Ce