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酿酒酵母ACR3基因编码一种参与亚砷酸盐转运的假定膜蛋白。

The Saccharomyces cerevisiae ACR3 gene encodes a putative membrane protein involved in arsenite transport.

作者信息

Wysocki R, Bobrowicz P, Ułaszewski S

机构信息

Institute of Microbiology, Wroclaw University, Przybyszewskiego 63/77, 51-148 Wroclaw, Poland.

出版信息

J Biol Chem. 1997 Nov 28;272(48):30061-6. doi: 10.1074/jbc.272.48.30061.

Abstract

The cluster of three genes, ACR1, ACR2, and ACR3, previously was shown to confer arsenical resistance in Saccharomyces cerevisiae. The overexpression of ACR3 induced high level arsenite resistance. The presence of ACR3 together with ACR2 on a multicopy plasmid was conducive to increased arsenate resistance. The function of ACR3 gene has now been investigated. Amino acid sequence analysis of Acr3p showed that this hypothetical protein has hydrophobic character with 10 putative transmembrane spans and is probably located in yeast plasma membrane. We constructed the acr3 null mutation. The resulting disruptants were 5-fold more sensitive to arsenate and arsenite than wild-type cells. The acr3 disruptants showed wild-type sensitivity to antimony, tellurite, cadmium, and phenylarsine oxide. The mechanism of arsenical resistance was assayed by transport experiments using radioactive arsenite. We did not observe any significant differences in the accumulation of 76AsO33- in wild-type cells, acr1 and acr3 disruptants. However, the high dosage of ACR3 gene resulted in loss of arsenite uptake. These results suggest that arsenite resistance in yeast is mediated by an arsenite transporter (Acr3p).

摘要

先前已证明,由ACR1、ACR2和ACR3这三个基因组成的基因簇可赋予酿酒酵母抗砷性。ACR3的过表达可诱导高水平的亚砷酸盐抗性。ACR3与ACR2一起存在于多拷贝质粒上有助于增强对砷酸盐的抗性。现在已经对ACR3基因的功能进行了研究。对Acr3p的氨基酸序列分析表明,这种假定的蛋白质具有疏水性,有10个推定的跨膜结构域,可能位于酵母质膜中。我们构建了acr3缺失突变体。所得的缺失突变体对砷酸盐和亚砷酸盐的敏感性比野生型细胞高5倍。acr3缺失突变体对锑、亚碲酸盐、镉和氧化苯胂表现出野生型敏感性。通过使用放射性亚砷酸盐的转运实验测定了抗砷机制。我们没有观察到野生型细胞、acr1和acr3缺失突变体中76AsO33-积累的任何显著差异。然而,高剂量的ACR3基因导致亚砷酸盐摄取丧失。这些结果表明,酵母中的亚砷酸盐抗性是由一种亚砷酸盐转运蛋白(Acr3p)介导的。

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