Dominant negative alleles of SEC10 reveal distinct domains involved in secretion and morphogenesis in yeast.

Dominant negative alleles of SEC10 reveal distinct domains involved in secretion and morphogenesis in yeast.
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SEC10 的显性负等位基因揭示了参与酵母分泌和形态发生的不同结构域。

DOI:
10.1091/mbc.9.7.1725
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发表时间:
1998
影响因子:
3.3
通讯作者:
Novick,P
Novick,P
中科院分区:
生物学3区
文献类型:
--
作者:
Roth,D;Guo,W;Novick,P

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将分泌囊泡精确靶向至质膜上的不同位点对于实现极化生长和在真核细胞表面建立专门的结构域是必要的。胞吐作用所需的蛋白质复合物的成员,即外囊,已经定位于芽殖酵母中的活性分泌区域,在那里它们可以发挥功能,指定质膜上用于囊泡对接和融合的位点。在这项研究中,我们已经解决了一个成员的外囊复合物,Sec10 p的功能。我们已经确定了Sec10p的两个功能域,它们以显性负性方式抑制过表达后的细胞生长。显性阴性突变体的表型和生化分析表明Sec10p具有双功能作用。其中一个结构域由Sec10 p氨基末端的三分之二组成,直接与Sec15 p(另一个外囊组分)相互作用。该结构域的过表达从胞吐复合物中置换全长Sec10,导致胞吐作用的阻断和分泌囊泡的积累。Sec10 p的羧基末端结构域不与外囊复合物的其他成员相互作用,并且该结构域的表达不引起分泌缺陷。相反,这种突变导致形成细长的细胞,这表明Sec10 p的第二个结构域是形态发生所需的,可能调节细胞周期进程中从新生子细胞尖端到母女连接的分泌途径的重新定位。
The accurate targeting of secretory vesicles to distinct sites on the plasma membrane is necessary to achieve polarized growth and to establish specialized domains at the surface of eukaryotic cells. Members of a protein complex required for exocytosis, the exocyst, have been localized to regions of active secretion in the budding yeastSaccharomyces cerevisiaewhere they may function to specify sites on the plasma membrane for vesicle docking and fusion. In this study we have addressed the function of one member of the exocyst complex, Sec10p. We have identified two functional domains of Sec10p that act in a dominant-negative manner to inhibit cell growth upon overexpression. Phenotypic and biochemical analysis of the dominant-negative mutants points to a bifunctional role for Sec10p. One domain, consisting of the amino-terminal two-thirds of Sec10p directly interacts with Sec15p, another exocyst component. Overexpression of this domain displaces the full-length Sec10 from the exocyst complex, resulting in a block in exocytosis and an accumulation of secretory vesicles. The carboxy-terminal domain of Sec10p does not interact with other members of the exocyst complex and expression of this domain does not cause a secretory defect. Rather, this mutant results in the formation of elongated cells, suggesting that the second domain of Sec10p is required for morphogenesis, perhaps regulating the reorientation of the secretory pathway from the tip of the emerging daughter cell toward the mother–daughter connection during cell cycle progression.
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