Salama N R, Chuang J S, Schekman R W
Department of Molecular and Cell Biology, Howard Hughes Research Institute, University of California, Berkeley 94720, USA.
Mol Biol Cell. 1997 Feb;8(2):205-17. doi: 10.1091/mbc.8.2.205.
The COPII vesicle coat protein promotes the formation of endoplasmic reticulum- (ER) derived transport vesicles that carry secretory proteins to the Golgi complex in Saccharomyces cerevisiae. This coat protein consists of Sar1p, the Sec23p protein complex containing Sec23p and Sec24p, and the Sec13p protein complex containing Sec13p and a novel 150-kDa protein, p150. Here, we report the cloning and characterization of the p150 gene. p150 is encoded by an essential gene. Depletion of this protein in vivo blocks the exit of secretory proteins from the ER and causes an elaboration of ER membranes, indicating that p150 is encoded by a SEC gene. Additionally, overproduction of the p150 gene product compromises the growth of two ER to Golgi sec mutants: sec16-2 and sec23-1. p150 is encoded by SEC31, a gene isolated in a genetic screen for mutations that accumulate unprocessed forms of the secretory protein alpha-factor. The sec31-1 mutation was mapped by gap repair, and sequence analysis revealed an alanine to valine change at position 1239, near the carboxyl terminus. Sec31p is a phosphoprotein and treatment of the Sec31p-containing fraction with alkaline phosphatase results in a 50-75% inhibition of transport vesicle formation activity in an ER membrane budding assay.
COPII囊泡外被蛋白促进酿酒酵母中内质网(ER)衍生的运输囊泡的形成,这些囊泡将分泌蛋白运输到高尔基体复合体。这种外被蛋白由Sar1p、包含Sec23p和Sec24p的Sec23p蛋白复合体以及包含Sec13p和一种新的150 kDa蛋白p150的Sec13p蛋白复合体组成。在此,我们报告了p150基因的克隆和特性分析。p150由一个必需基因编码。体内该蛋白的缺失会阻止分泌蛋白从内质网输出,并导致内质网膜的扩张,这表明p150由一个SEC基因编码。此外,p150基因产物的过量表达会损害两个内质网到高尔基体的sec突变体sec16 - 2和sec23 - 1的生长。p150由SEC31编码,SEC31是在一个针对积累未加工形式分泌蛋白α因子的突变体的遗传筛选中分离出的基因。通过缺口修复对sec31 - 1突变进行定位,序列分析显示在靠近羧基末端的第(原文此处有误,结合后文推测为1239)位发生了丙氨酸到缬氨酸的变化。Sec31p是一种磷蛋白,在用碱性磷酸酶处理含有Sec31p的组分后,在内质网膜出芽试验中运输囊泡形成活性受到50 - 75%的抑制。