Institut für Zelluläre und Molekulare Botanik, Universität Bon, D-53115 Bon, Germany.
Plant Cell. 2009 Dec;21(12):4018-30. doi: 10.1105/tpc.109.070557. Epub 2009 Dec 4.
The MRS2/MGT gene family in Arabidopsis thaliana belongs to the superfamily of CorA-MRS2-ALR-type membrane proteins. Proteins of this type are characterized by a GMN tripeptide motif (Gly-Met-Asn) at the end of the first of two C-terminal transmembrane domains and have been characterized as magnesium transporters. Using the recently established mag-fura-2 system allowing direct measurement of Mg(2+) uptake into mitochondria of Saccharomyces cerevisiae, we find that all members of the Arabidopsis family complement the corresponding yeast mrs2 mutant. Highly different patterns of tissue-specific expression were observed for the MRS2/MGT family members in planta. Six of them are expressed in root tissues, indicating a possible involvement in plant magnesium supply and distribution after uptake from the soil substrate. Homozygous T-DNA insertion knockout lines were obtained for four members of the MRS2/MGT gene family. A strong, magnesium-dependent phenotype of growth retardation was found for mrs2-7 when Mg(2+) concentrations were lowered to 50 microM in hydroponic cultures. Ectopic overexpression of MRS2-7 from the cauliflower mosaic virus 35S promoter results in complementation and increased biomass accumulation. Green fluorescent protein reporter gene fusions indicate a location of MRS2-7 in the endomembrane system. Hence, contrary to what is frequently found in analyses of plant gene families, a single gene family member knockout results in a strong, environmentally dependent phenotype.
拟南芥 MRS2/MGT 基因家族属于 CorA-MRS2-ALR 型膜蛋白超家族。这类蛋白的特征是在两个 C 端跨膜结构域的第一个末端有 GMN 三肽基序(Gly-Met-Asn),并被表征为镁转运蛋白。利用最近建立的 mag-fura-2 系统,可以直接测量 Saccharomyces cerevisiae 线粒体中镁的摄取,我们发现拟南芥家族的所有成员都能互补相应的酵母 mrs2 突变体。在植物中,MRS2/MGT 家族成员的组织特异性表达呈现出高度不同的模式。其中 6 种在根组织中表达,表明它们可能参与了植物从土壤基质中吸收后镁的供应和分布。我们获得了 MRS2/MGT 基因家族四个成员的纯合 T-DNA 插入敲除系。当在水培培养物中将镁浓度降低至 50 μM 时,mrs2-7 表现出强烈的、依赖于镁的生长迟缓表型。来自花椰菜花叶病毒 35S 启动子的 MRS2-7 的异位过表达导致互补和生物量积累增加。绿色荧光蛋白报告基因融合表明 MRS2-7 位于内膜系统中。因此,与植物基因家族分析中经常发现的情况相反,单个基因家族成员的敲除会导致强烈的、依赖于环境的表型。