Institut des Sciences de la Vie, Université Catholique de Louvain, Croix du Sud 4-15, B-1348 Louvain-la-Neuve, Belgium.
Plant J. 2011 Apr;66(2):306-17. doi: 10.1111/j.1365-313X.2011.04496.x. Epub 2011 Mar 1.
Major intrinsic proteins (MIPs) transport water and uncharged solutes across membranes in all kingdoms of life. Recently, an uncharacterized MIP subfamily was identified in the genomes of plants and fungi and named X Intrinsic Proteins (XIPs). Here, we describe the genetic features, localization, expression, and functions of a group of Solanaceae XIPs. XIP cDNA and gDNA were cloned from tobacco, potato, tomato, and morning glory. A conserved sequence motif in the first intron of Solanaceae XIPs initiates an RNA-processing mechanism that results in two splice variants (α and β). When transiently or stably expressed in tobacco plants, yellow fluorescent protein-tagged NtXIP1;1α and NtXIP1;1β were both localized in the plasma membrane. Transgenic tobacco lines expressing NtXIP1;1-promoter-GUS constructs and RT-PCR studies showed that NtXIP1;1 was expressed in all organs. The NtXIP1;1 promoter was mainly active in cell layers facing the environment in all above-ground tissues. Heterologous expression of Solanaceae XIPs in Xenopus laevis oocytes and various Saccharomyces cerevisiae mutants demonstrated that these isoforms facilitate the transport of bulky solutes, such as glycerol, urea, and boric acid. In contrast, permeability for water was undetectable. These data suggest that XIPs function in the transport of uncharged solutes across the cell plasma membrane in specific plant tissues, including at the interface between the environment and external cell layers.
主要内在蛋白(MIPs)在所有生命领域的膜中运输水和不带电荷的溶质。最近,在植物和真菌的基因组中鉴定出一种未表征的 MIP 亚家族,并将其命名为 X 内在蛋白(XIPs)。在这里,我们描述了一组茄科 XIP 的遗传特征、定位、表达和功能。从烟草、马铃薯、番茄和牵牛花中克隆了 XIP cDNA 和 gDNA。茄科 XIP 第一内含子中的保守序列基序启动了一种 RNA 加工机制,导致两种剪接变体(α和β)。当瞬时或稳定表达在烟草植物中时,黄色荧光蛋白标记的 NtXIP1;1α 和 NtXIP1;1β 均定位于质膜。表达 NtXIP1;1 启动子-GUS 构建体的转基因烟草系和 RT-PCR 研究表明,NtXIP1;1 在所有器官中均有表达。NtXIP1;1 启动子主要在所有地上组织中面向环境的细胞层中活跃。茄科 XIPs 在非洲爪蟾卵母细胞和各种酿酒酵母突变体中的异源表达表明,这些同工型促进了甘油、尿素和硼酸等大溶质的运输。相比之下,水的通透性无法检测到。这些数据表明,XIPs 在特定植物组织的细胞质膜中运输不带电荷的溶质中起作用,包括在环境和外部细胞层之间的界面处。