Clemens S, Antosiewicz D M, Ward J M, Schachtman D P, Schroeder J I
Department of Biology and Center for Molecular Genetics, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0116, USA.
Proc Natl Acad Sci U S A. 1998 Sep 29;95(20):12043-8. doi: 10.1073/pnas.95.20.12043.
Nonessential metal ions such as cadmium are most likely transported across plant membranes via transporters for essential cations. To identify possible pathways for Cd2+ transport we tested putative plant cation transporters for Cd2+ uptake activity by expressing cDNAs in Saccharomyces cerevisiae and found that expression of one clone, LCT1, renders the growth of yeast more sensitive to cadmium. Ion flux assays showed that Cd2+ sensitivity is correlated with an increase in Cd2+ uptake. LCT1-dependent Cd2+ uptake is saturable, lies in the high-affinity range (apparent KM for Cd2+ = 33 microM) and is sensitive to block by La3+ and Ca2+. Growth assays demonstrated a sensitivity of LCT1-expressing yeast cells to extracellular millimolar Ca2+ concentrations. LCT1-dependent increase in Ca2+ uptake correlated with the observed phenotype. Furthermore, LCT1 complements a yeast disruption mutant in the MID1 gene, a non-LCT1-homologous yeast gene encoding a membrane Ca2+ influx system required for recovery from the mating response. We conclude that LCT1 mediates the uptake of Ca2+ and Cd2+ in yeast and may therefore represent a first plant cDNA encoding a plant Ca2+ uptake or an organellar Ca2+ transport pathway in plants and may contribute to transport of the toxic metal Cd2+ across plant membranes.
诸如镉等非必需金属离子很可能是通过必需阳离子转运蛋白跨植物细胞膜运输的。为了确定Cd2+运输的可能途径,我们通过在酿酒酵母中表达cDNA来测试假定的植物阳离子转运蛋白的Cd2+摄取活性,发现一个克隆LCT1的表达使酵母的生长对镉更敏感。离子通量分析表明,Cd2+敏感性与Cd2+摄取的增加相关。依赖LCT1的Cd2+摄取是可饱和的,处于高亲和力范围内(Cd2+的表观KM = 33 microM),并且对La3+和Ca2+的阻断敏感。生长分析表明,表达LCT1的酵母细胞对细胞外毫摩尔浓度的Ca2+敏感。依赖LCT1的Ca2+摄取增加与观察到的表型相关。此外,LCT1补充了MID1基因中的酵母破坏突变体,MID1是一个与LCT1非同源的酵母基因,编码从交配反应中恢复所需的膜Ca2+内流系统。我们得出结论,LCT1介导酵母中Ca2+和Cd2+的摄取,因此可能代表第一个编码植物Ca2+摄取或植物细胞器Ca2+运输途径的植物cDNA,并且可能有助于有毒金属Cd2+跨植物细胞膜的运输。