Molecular cloning and expression of the Saccharomyces cerevisiae STS1 gene product. A yeast ABC transporter conferring mycotoxin resistance.
Molecular cloning and expression of the Saccharomyces cerevisiae STS1 gene product. A yeast ABC transporter conferring mycotoxin resistance.
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DOI：
10.1016/s0021-9258(17)41760-1
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发表时间：
1994-02
期刊：
影响因子：
--
通讯作者：
P. Bissinger;K. Kuchler
中科院分区：
文献类型：
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作者：
P. Bissinger;K. Kuchler
我们克隆了一种酵母基因，当存在于多个拷贝时，该基因赋予酿酒酵母多药耐药表型。STS1 （Sporidesmin Toxicity Suppressor）基因编码一个1511-残基蛋白，其预测的结构组织具有12个α -螺旋膜片段和两个包含ATP结合一致位点的结构域，表明STS1是一种新的酵母ATP结合盒（ABC）转运体。染色体上STS1的缺失会导致两种交配类型的δ STS1细胞存活，这表明STS1对细胞生长不是必需的。然而，三角洲sts1细胞对孢子素和其他结构无关的药物如环己亚胺表现出超敏感性。相反，STS1的过表达会导致对相同药物的耐药性增加。尽管Northern分析表明STS1 mRNA存在于所有酵母细胞类型中，但其在α因子阻滞细胞中的水平急剧降低表明STS1的表达受交配信息素的调节。利用单克隆抗体进行亚细胞分离和免疫印迹，发现Sts1是一个175 kda的膜蛋白，主要定位于细胞膜内。
We have cloned a yeast gene that confers a multidrug resistance phenotype on Saccharomyces cerevisiae when present in multiple copies. The STS1 (for Sporidesmin Toxicity Suppressor) gene encodes a 1511-residue protein whose predicted structural organization is characterized by 12 alpha-helical membrane segments and two domains containing consensus sites for ATP binding, indicating that STS1 is a new yeast ATP-binding cassette (ABC) transporter. A chromosomal deletion of STS1 leads to viable delta sts1 cells of both mating types, suggesting that STS1 is not essential for cell growth. However, delta sts1 cells exhibit supersensitivity to sporidesmin and to other structurally unrelated drugs such as cycloheximide. Conversely, overexpression of STS1 leads to increased resistance to the same drugs. Although Northern analysis showed that STS1 mRNA is present in all yeast cell types, its drastically reduced level in alpha-factor-arrested cells indicates that expression of STS1 is regulated by mating pheromones. Subcellular fractionation and immunoblotting using monoclonal antibodies, which recognize a fully functional epitope-tagged Sts1 protein, showed that Sts1 is a 175-kDa membrane protein localized mainly to intracellular membranes.