Etheridge N, Trusov Y, Verbelen J P, Botella J R
Department of Botany, University of Queensland, Brisbane, Australia.
Plant Mol Biol. 1999 Apr;39(6):1113-26. doi: 10.1023/a:1006137221259.
We report the initial characterization of an Arabidopsis thaliana cDNA (atdrg1), a member of a new class of GTP-binding proteins (G-proteins) in plants. The predicted ATDRG1 protein contains all five structural motifs characteristic of the G-protein superfamily. Apart from these motifs, the amino acid sequence differs substantially from all known G-proteins except for a recently discovered new family named developmentally regulated G-proteins (DRGs). Sequences closely related to atdrg1 are found in species as distant as human (80% amino acid conservation), Drosophila (74%), yeast (77%) and Caenorhabditis elegans (77%). The remarkable evolutionary conservation of these proteins suggests an important, but as yet unclear role. Phylogenetic analysis of the available homologous sequences strongly suggests a diphyletic origin of the eukaryotic DRG proteins. Northern analysis shows high levels of atdrg1 mRNA in all Arabidopsis tissues studied, and homologues of atdrg1 are present throughout the plant kingdom. In situ hybridization reveals that atdrg1 is highly expressed in actively growing tissues and reproductive organs. Southern analysis indicates the presence of either one or two copies of atdrg1 in the Arabidopsis genome. Immunolocalization studies show that the protein is present in cytoplasmic vesicles found mainly in actively growing tissues suggesting a putative role for ATDRG1 in either the regulation of vesicle transport or the regulation of enzymes involved in storage protein processing.
我们报道了拟南芥cDNA(atdrg1)的初步特征,它是植物中一类新的GTP结合蛋白(G蛋白)的成员。预测的ATDRG1蛋白包含G蛋白超家族所有五个特征性结构基序。除了这些基序外,其氨基酸序列与所有已知的G蛋白有很大差异,除了最近发现的一个名为发育调控G蛋白(DRG)的新家族。在与拟南芥亲缘关系甚远的物种中,如人类(氨基酸保守率80%)、果蝇(74%)、酵母(77%)和秀丽隐杆线虫(77%),都发现了与atdrg1密切相关的序列。这些蛋白显著的进化保守性表明其具有重要但尚未明确的作用。对现有同源序列的系统发育分析强烈表明真核DRG蛋白有双系起源。Northern分析显示,在所有研究的拟南芥组织中atdrg1 mRNA水平都很高,并且atdrg1的同源物存在于整个植物界。原位杂交显示atdrg1在活跃生长的组织和生殖器官中高度表达。Southern分析表明拟南芥基因组中存在一个或两个atdrg1拷贝。免疫定位研究表明,该蛋白存在于主要在活跃生长组织中发现的细胞质小泡中,这表明ATDRG1在囊泡运输调节或参与储存蛋白加工的酶的调节中可能发挥作用。