Division of Biological Sciences, Cell and Developmental Biology Section, University of California, San Diego, La Jolla, California 92093, USA.
J Biol Chem. 2010 Dec 24;285(52):40416-26. doi: 10.1074/jbc.M110.155408. Epub 2010 Oct 11.
Phytochelatins mediate tolerance to heavy metals in plants and some fungi by sequestering phytochelatin-metal complexes into vacuoles. To date, only Schizosaccharomyces pombe Hmt1 has been described as a phytochelatin transporter and attempts to identify orthologous phytochelatin transporters in plants and other organisms have failed. Furthermore, recent data indicate that the hmt1 mutant accumulates significant phytochelatin levels in vacuoles, suggesting that unidentified phytochelatin transporters exist in fungi. Here, we show that deletion of all vacuolar ABC transporters abolishes phytochelatin accumulation in S. pombe vacuoles and abrogates (35)S-PC(2) uptake into S. pombe microsomal vesicles. Systematic analysis of the entire S. pombe ABC transporter family identified Abc2 as a full-size ABC transporter (ABCC-type) that mediates phytochelatin transport into vacuoles. The S. pombe abc1 abc2 abc3 abc4 hmt1 quintuple and abc2 hmt1 double mutant show no detectable phytochelatins in vacuoles. Abc2 expression restores phytochelatin accumulation into vacuoles and suppresses the cadmium sensitivity of the abc quintuple mutant. A novel, unexpected, function of Hmt1 in GS-conjugate transport is also shown. In contrast to Hmt1, Abc2 orthologs are widely distributed among kingdoms and are proposed as the long-sought vacuolar phytochelatin transporters in plants and other organisms.
植物螯合肽通过将植物螯合肽-金属复合物螯合到液泡中来介导植物和一些真菌对重金属的耐受。迄今为止,只有酿酒酵母 Hmt1 被描述为植物螯合肽转运蛋白,并且试图在植物和其他生物体中鉴定同源植物螯合肽转运蛋白的尝试都失败了。此外,最近的数据表明,hmt1 突变体在液泡中积累了大量的植物螯合肽,这表明真菌中存在未鉴定的植物螯合肽转运蛋白。在这里,我们表明,所有液泡 ABC 转运蛋白的缺失都会导致酿酒酵母液泡中植物螯合肽的积累,并消除(35)S-PC(2)进入酿酒酵母微粒体囊泡的摄取。对整个酿酒酵母 ABC 转运蛋白家族的系统分析鉴定出 Abc2 为一种全尺寸 ABC 转运蛋白(ABCC 型),可将植物螯合肽转运到液泡中。酿酒酵母 abc1 abc2 abc3 abc4 hmt1 五突变体和 abc2 hmt1 双突变体在液泡中检测不到可检测到的植物螯合肽。Abc2 表达恢复了植物螯合肽在液泡中的积累,并抑制了 abc 五突变体对镉的敏感性。还显示了 Hmt1 在 GS 缀合物转运中的一个新的、意想不到的功能。与 Hmt1 相反,Abc2 同源物广泛分布于各个王国,被提议作为植物和其他生物体中寻找已久的液泡植物螯合肽转运蛋白。