Graduate School of Life Sciences, Tohoku University, 2-1-1 Katahira, Aoba, Sendai, Miyagi 980-8577, Japan.
J Exp Bot. 2013 Nov;64(14):4517-27. doi: 10.1093/jxb/ert267.
A rice cDNA, OsDEP1, encoding a highly cysteine (Cys)-rich G protein γ subunit, was initially identified as it conferred cadmium (Cd) tolerance on yeast cells. Of the 426 aa constituting OsDEP1, 120 are Cys residues (28.2%), of which 88 are clustered in the C-terminal half region (aa 170-426). To evaluate the independent effects of these two regions, two truncated versions of the OsDEP1-expressing plasmids pOsDEP1(1-169) and pOsDEP1(170-426) were used to examine their effects on yeast Cd tolerance. Although OsDEP1(170-426) conferred a similar level of Cd tolerance as the intact OsDEP1, OsDEP1(1-169) provided no such tolerance, indicating that the tolerance effect is localized to the aa 170-426 C-terminal peptide region. The Cd responses of transgenic Arabidopsis plants constitutively expressing OsDEP1, OsDEP1(1-169) or OsDEP1(170-426), were similar to the observations in yeast cells, with OsDEP1 and OsDEP1(170-426) transgenic plants displaying Cd tolerance but OsDEP1(1-169) plants showing no such tolerance. In addition, a positive correlation between the transcript levels of OsDEP1 or OsDEP1(170-426) in the transgenics and the Cd content of these plants upon Cd application was observed. As several Arabidopsis loss-of-function heterotrimeric G protein β and γ subunit gene mutants did not show differences in their Cd sensitivity compared with wild-type plants, we propose that the Cys-rich region of OsDEP1 may function directly as a trap for Cd ions.
一个水稻 cDNA,OsDEP1,编码一个高度富含半胱氨酸(Cys)的 G 蛋白γ亚基,最初被鉴定为它赋予酵母细胞镉(Cd)耐受性。在构成 OsDEP1 的 426 个氨基酸中,有 120 个是半胱氨酸残基(28.2%),其中 88 个聚集在 C 末端半区(氨基酸 170-426)。为了评估这两个区域的独立效应,使用两个截断的 OsDEP1 表达质粒 pOsDEP1(1-169)和 pOsDEP1(170-426)来检验它们对酵母 Cd 耐受性的影响。虽然 OsDEP1(170-426)赋予了与完整 OsDEP1 相似的 Cd 耐受性,但 OsDEP1(1-169)没有提供这种耐受性,表明这种耐受性效应定位于 aa 170-426 C 末端肽区。持续表达 OsDEP1、OsDEP1(1-169)或 OsDEP1(170-426)的转基因拟南芥植物的 Cd 反应与酵母细胞的观察结果相似,OsDEP1 和 OsDEP1(170-426)转基因植物表现出 Cd 耐受性,但 OsDEP1(1-169)植物没有表现出这种耐受性。此外,在 Cd 处理后,转基因植物中 OsDEP1 或 OsDEP1(170-426)的转录水平与这些植物的 Cd 含量之间存在正相关。由于几个拟南芥失活的异三聚体 G 蛋白β和γ亚基基因突变体与野生型植物相比在 Cd 敏感性方面没有差异,我们提出 OsDEP1 的富含半胱氨酸区可能直接作为 Cd 离子的陷阱发挥作用。