An aquaporin PvTIP4;1 from Pteris vittata may mediate arsenite uptake
An aquaporin PvTIP4;1 from Pteris vittata may mediate arsenite uptake
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来自 Pteris vittata 的水通道蛋白 PvTIP4;1 可能介导亚砷酸盐的吸收
DOI：
10.1111/nph.13637
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发表时间：
2016-01-01
期刊：
影响因子：
9.4
通讯作者：
Ma, Mi
中科院分区：
文献类型：
--
作者：
He, Zhenyan;Yan, Huili;Ma, Mi
蕨类植物狗牙蕨是砷超富集植物。参与亚砷酸盐（As（III））运输的基因尚不清楚。本文报道了一个新的可能介导As（III）摄取的沙芥水通道蛋白基因PvTIP 4; 1的分离和鉴定。在酿酒酵母和拟南芥中分析了PvTIP 4;1的砷毒性和积累活性。对PvTIP 4;1-GFP融合蛋白在杜仲原生质体和愈伤组织中的亚细胞定位进行了研究。采用实时荧光定量PCR检测PvTIP 4; 1的组织表达。研究了PvTIP 4;1芳香/精氨酸（Ar/R）结构域的定点突变，在酵母中的异源表达表明PvTIP 4;1能够促进As（III）的扩散。转基因拟南芥表明，PvTIP 4;1增加砷的积累和诱导砷敏感性。图像和FM 4 -64染色表明，PvTIP 4;1定位于P. vittata细胞的质膜。组织定位研究表明，PvTIP 4;1转录本主要在根中表达。PvTIP 4;1是一个新的代表性液泡膜内在蛋白（TIP）水通道蛋白，其功能和定位结果表明PvTIP 4;1可能参与了对As（III）的摄取。
The fern Pteris vittata is an arsenic hyperaccumulator. The genes involved in arsenite (As (III)) transport are not yet clear. Here, we describe the isolation and characterization of a new P. vittata aquaporin gene, PvTIP4;1, which may mediate As(III) uptake.PvTIP4;1 was identified from yeast functional complement cDNA library of P. vittata. Arsenic toxicity and accumulating activities of PvTIP4;1 were analyzed in Saccharomyces cerevisiae and Arabidopsis. Subcellular localization of PvTIP4;1-GFP fusion protein in P. vittata protoplast and callus was conducted. The tissue expression of PvTIP4; 1 was investigated by quantitative real-time PCR. Site-directed mutagenesis of the PvTIP4;1 aromatic/arginine (Ar/R) domain was studied.Heterologous expression in yeast demonstrates that PvTIP4;1 was able to facilitate As(III) diffusion. Transgenic Arabidopsis showed that PvTIP4;1 increases arsenic accumulation and induces arsenic sensitivity. Images and FM4-64 staining suggest that PvTIP4;1 localizes to the plasma membrane in P. vittata cells. A tissue location study shows that PvTIP4;1 transcripts are mainly expressed in roots. Site-directed mutation in yeast further proved that the cysteine at the LE1 position of PvTIP4;1 Ar/R domain is a functional site.PvTIP4;1 is a new represented tonoplast intrinsic protein (TIP) aquaporin from P. vittata and the function and location results imply that PvTIP4;1 may be involved in As(III) uptake.