An arf1Delta synthetic lethal screen identifies a new clathrin heavy chain conditional allele that perturbs vacuolar protein transport in Saccharomyces cerevisiae.
An arf1Delta synthetic lethal screen identifies a new clathrin heavy chain conditional allele that perturbs vacuolar protein transport in Saccharomyces cerevisiae.
复制标题
arf1Delta 合成致死筛选鉴定出一种新的网格蛋白重链条件等位基因，该等位基因扰乱酿酒酵母中的液泡蛋白运输。
DOI：
10.1093/genetics/150.2.577
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发表时间：
1998
期刊：
影响因子：
3.3
通讯作者：
Graham,TR
中科院分区：
文献类型：
--
作者：
Chen,CY;Graham,TR
ADP-核糖化因子(ARF)是一种小的GTP结合蛋白，被认为调节外壳蛋白在运输小泡上的组装。为了确定与ARF功能相互作用的因素，我们在酿酒酵母中进行了基因筛查，以寻找具有ARF1Δ等位基因的合成致死性突变，并通过互补测试定义了七个基因(SWA1-7用于合成LETHARF1Δ)。大多数突变体表现出与ARF1Δ突变体相似的表型，如温度条件生长，对氟离子超敏，以及部分蛋白质运输和糖基化缺陷。在这里，我们报道了SWA5-1是一种新的温度敏感等位基因，它携带着CHC1开放阅读框3‘端附近的移码突变。Arf1和chc1之间的这种遗传相互作用为ARF在笼状蛋白被层组装中的作用提供了体内证据。令人惊讶的是，含有chc1-5的菌株在将羧基肽酶Y或羧基肽酶S运输到液泡的过程中表现出显著的缺陷，这在其他chc1-ts突变体中没有观察到。在chc1-5突变体中，转移酶的分泌或碱性磷酸酶向液泡的运输动力学没有受到显著影响，进一步暗示了Cathrin在高尔基体到液泡的运输途径中具有特异性。
ADP-ribosylation factor (ARF) is a small GTP-binding protein that is thought to regulate the assembly of coat proteins on transport vesicles. To identify factors that functionally interact with ARF, we have performed a genetic screen in Saccharomyces cerevisiae for mutations that exhibit synthetic lethality with anarf1Δ allele and defined seven genes by complementation tests (SWA1-7 forsynthetically lethalwitharf1Δ). Most of the swa mutants exhibit phenotypes comparable to arf1Δ mutants such as temperature-conditional growth, hypersensitivity to fluoride ions, and partial protein transport and glycosylation defects. Here, we report that swa5-1 is a new temperature-sensitive allele of the clathrin heavy chain gene (chc1-5), which carries a frameshift mutation near the 3′ end of the CHC1 open reading frame. This genetic interaction between arf1 and chc1 provides in vivo evidence for a role for ARF in clathrin coat assembly. Surprisingly, strains harboring chc1-5 exhibited a significant defect in transport of carboxypeptidase Y or carboxypeptidase S to the vacuole that was not observed in other chc1 ts mutants. The kinetics of invertase secretion or transport of alkaline phosphatase to the vacuole were not significantly affected in the chc1-5 mutant, further implicating clathrin specifically in the Golgi to vacuole transport pathway for carboxypeptidase Y.