Weig A, Deswarte C, Chrispeels M J
Department of Biology, University of California, San Diego, La Jolla 92093-0116, USA.
Plant Physiol. 1997 Aug;114(4):1347-57. doi: 10.1104/pp.114.4.1347.
Aquaporins, proteins that enhance the permeability of biological membranes to water, are widely distributed in living organisms. They are 26- to 29-kD proteins that belong to the major intrinsic protein (MIP) family of channels. By searching the Arabidopsis thaliana expressed sequence tag database and by using the polymerase chain reaction with oligonucleotides to conserved plant aquaporin domains, we identified 23 expressed Arabidopsis MIP genes. Eight of these had been previously identified as active aquaporins, and two additional ones are now reported to have water-transport activity in Xenopus laevis oocytes. One of these is highly expressed in suspension-cultured cells. On a dendrogram these 23 MIP sequences cluster into three groups: the first group has 11 members and contains the plasma membrane aquaporins, the second group also has 11 members and contains the tonoplast aquaporins, and the third group has only a single member. This MIP protein, provisionally called At-NLM1, is most closely related to the Gm-NOD26 protein that is found in the bacteroid membranes of soybean (Glycine max L.) nodules; At-NLM1 is an active aquaporin when expressed in oocytes. With a semiquantitative slot-blot analysis technique, we determined the expression levels of 22 MIP genes in the various organs. The quantitative polymerase chain reaction was used to determine the effects of various stress treatments on the expression of NLM1.
水通道蛋白是一类能提高生物膜对水通透性的蛋白质,广泛分布于生物体内。它们是26至29千道尔顿的蛋白质,属于主要内在蛋白(MIP)通道家族。通过搜索拟南芥表达序列标签数据库,并使用针对保守植物水通道蛋白结构域的寡核苷酸进行聚合酶链反应,我们鉴定出23个已表达的拟南芥MIP基因。其中8个先前已被鉴定为活性水通道蛋白,现在又有另外两个被报道在非洲爪蟾卵母细胞中具有水运输活性。其中一个在悬浮培养细胞中高度表达。在一个系统发育树上,这23个MIP序列聚为三组:第一组有11个成员,包含质膜水通道蛋白;第二组也有11个成员,包含液泡膜水通道蛋白;第三组只有一个成员。这种MIP蛋白暂称为At-NLM1,与在大豆(Glycine max L.)根瘤类菌体膜中发现的Gm-NOD26蛋白关系最为密切;At-NLM1在卵母细胞中表达时是一种活性水通道蛋白。通过半定量狭缝印迹分析技术,我们测定了22个MIP基因在各个器官中的表达水平。使用定量聚合酶链反应来确定各种胁迫处理对NLM1表达的影响。