The Exocyst is a multiprotein complex required for exocytosis in Saccharomyces cerevisiae
The Exocyst is a multiprotein complex required for exocytosis in Saccharomyces cerevisiae
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DOI：
10.1002/j.1460-2075.1996.tb01039.x
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发表时间：
1996-12-02
期刊：
影响因子：
11.4
通讯作者：
Novick, P
中科院分区：
文献类型：
--
作者：
TerBush, DR;Maurice, T;Novick, P
在酿酒酵母中，至少有15个基因的产物特异性地参与囊泡从高尔基器到质膜的运输，此前，我们已经证明其中的三个基因SEC6、SEC8和SEC15编码位于芽尖的多亚单位复合体的组成部分，芽尖是酿酒酵母胞吐的主要部位。另外三个基因SEC3、SEC5和SEC10的突变被发现破坏了Sec6-Sec8-Sec15复合体的亚基完整性，表明这些基因可能编码这个复合体的一些剩余成分。为了验证这种可能性，我们克隆并测序了Secs和SEC10基因，破坏了它们，并将其标记(Sec5p)或制备了多克隆抗血清(Sec10p)用于免疫共沉淀研究。同时，我们生化提纯了Sec6-Sec8-Sec15复合体中剩余的未知多肽用于肽微测序，通过预测的氨基酸序列与纯化的复合体组分的肽微测序结果进行比较，确定了编码这些组分的基因。除了Sec6p、Sec8p和Sec15p外，该复合体还包含由SEC3、Secs、SEC10和一个新基因Exo70编码的蛋白质，因为这七种蛋白共同作用于一个复合体中，而不是其他细胞内运输步骤，所以我们将其命名为Excost。
In the yeast Saccharomyces cerevisiae, the products of at least 15 genes are involved specifically in vesicular transport from the Golgi apparatus to the plasma membrane, Previously, we have shown that three of these genes, SEC6, SEC8 and SEC15, encode components of a multisubunit complex which localizes to the tip of the bud, the predominant site of exocytosis in S. cerevisiae. Mutations in three more of these genes, SEC3, SEC5 and SEC10, were found to disrupt the subunit integrity of the Sec6-Sec8-Sec15 complex, indicating that these genes may encode some of the remaining components of this complex, To examine this possibility, we cloned and sequenced the SECS and SEC10 genes, disrupted them, and either epitope tagged them (Sec5p) or prepared polyclonal antisera (Sec10p) to them for co-immunoprecipitation studies. Concurrently, we biochemically purified the remaining unidentified polypeptides of the Sec6-Sec8-Sec15 complex for peptide microsequencing, The genes encoding these components were identified by comparison of predicted amino acid sequences with those obtained from peptide microsequencing of the purified complex components, In addition to Sec6p, Sec8p and Sec15p, the complex contains the proteins encoded by SEC3, SECS, SEC10 and a novel gene, EXO70, Since these seven proteins function together in a complex required for exocytosis, and not other intracellular trafficking steps, we have named it the Exocyst.