Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, IL 60637 Department of Molecular Biology, Princeton University, Princeton, NJ 08544.
Mol Biol Cell. 2013 Nov;24(21):3406-19. doi: 10.1091/mbc.E13-04-0185. Epub 2013 Sep 4.
During the budding of coat protein complex II (COPII) vesicles from transitional endoplasmic reticulum (tER) sites, Sec16 has been proposed to play two distinct roles: negatively regulating COPII turnover and organizing COPII assembly at tER sites. We tested these ideas using the yeast Pichia pastoris. Redistribution of Sec16 to the cytosol accelerates tER dynamics, supporting a negative regulatory role for Sec16. To evaluate a possible COPII organization role, we dissected the functional regions of Sec16. The central conserved domain, which had been implicated in coordinating COPII assembly, is actually dispensable for normal tER structure. An upstream conserved region (UCR) localizes Sec16 to tER sites. The UCR binds COPII components, and removal of COPII from tER sites also removes Sec16, indicating that COPII recruits Sec16 rather than the other way around. We propose that Sec16 does not in fact organize COPII. Instead, regulation of COPII turnover can account for the influence of Sec16 on tER sites.
在衣壳蛋白复合物 II (COPII) 从过渡内质网 (tER) 部位出芽的过程中,Sec16 被提议发挥两个不同的作用:负调控 COPII 的周转率和在 tER 部位组织 COPII 的组装。我们使用酵母毕赤酵母测试了这些想法。Sec16 向细胞质的重新分配加速了 tER 的动态变化,支持 Sec16 的负调节作用。为了评估 COPII 组织作用的可能性,我们对 Sec16 的功能区域进行了剖析。中央保守结构域曾被认为参与协调 COPII 的组装,但实际上对于正常的 tER 结构是可有可无的。上游保守结构域 (UCR) 将 Sec16 定位在 tER 部位。UCR 与 COPII 成分结合,从 tER 部位去除 COPII 也会去除 Sec16,这表明 COPII 招募 Sec16,而不是相反。我们提出,Sec16 实际上并没有组织 COPII。相反,COPII 周转率的调节可以解释 Sec16 对 tER 部位的影响。