Rossanese O W, Soderholm J, Bevis B J, Sears I B, O'Connor J, Williamson E K, Glick B S
Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, Illinois 60637, USA.
J Cell Biol. 1999 Apr 5;145(1):69-81. doi: 10.1083/jcb.145.1.69.
Golgi stacks are often located near sites of "transitional ER" (tER), where COPII transport vesicles are produced. This juxtaposition may indicate that Golgi cisternae form at tER sites. To explore this idea, we examined two budding yeasts: Pichia pastoris, which has coherent Golgi stacks, and Saccharomyces cerevisiae, which has a dispersed Golgi. tER structures in the two yeasts were visualized using fusions between green fluorescent protein and COPII coat proteins. We also determined the localization of Sec12p, an ER membrane protein that initiates the COPII vesicle assembly pathway. In P. pastoris, Golgi stacks are adjacent to discrete tER sites that contain COPII coat proteins as well as Sec12p. This arrangement of the tER-Golgi system is independent of microtubules. In S. cerevisiae, COPII vesicles appear to be present throughout the cytoplasm and Sec12p is distributed throughout the ER, indicating that COPII vesicles bud from the entire ER network. We propose that P. pastoris has discrete tER sites and therefore generates coherent Golgi stacks, whereas S. cerevisiae has a delocalized tER and therefore generates a dispersed Golgi. These findings open the way for a molecular genetic analysis of tER sites.
高尔基体堆叠通常位于“过渡性内质网”(tER)附近,COPII转运囊泡在此产生。这种并列关系可能表明高尔基体潴泡在tER位点形成。为了探究这一观点,我们研究了两种出芽酵母:具有连贯高尔基体堆叠的巴斯德毕赤酵母和高尔基体分散的酿酒酵母。利用绿色荧光蛋白与COPII衣被蛋白的融合蛋白来观察这两种酵母中的tER结构。我们还确定了Sec12p的定位,Sec12p是一种内质网膜蛋白,它启动COPII囊泡组装途径。在巴斯德毕赤酵母中,高尔基体堆叠与离散的tER位点相邻,这些位点含有COPII衣被蛋白以及Sec12p。tER - 高尔基体系统的这种排列与微管无关。在酿酒酵母中,COPII囊泡似乎遍布整个细胞质,Sec12p分布在内质网各处,这表明COPII囊泡从整个内质网网络出芽。我们提出,巴斯德毕赤酵母具有离散的tER位点,因此产生连贯的高尔基体堆叠,而酿酒酵母具有分散的tER,因此产生分散的高尔基体。这些发现为tER位点的分子遗传学分析开辟了道路。