Teakle Graham R, Manfield Iain W, Graham John F, Gilmartin Philip M
Centre for Plant Sciences, University of Leeds, UK.
Plant Mol Biol. 2002 Sep;50(1):43-57. doi: 10.1023/a:1016062325584.
Many light-responsive promoters contain GATA motifs and a number of nuclear proteins have been defined that interact with these elements. Type-IV zinc-finger proteins have been extensively characterised in animals and fungi and are referred to as GATA factors by virtue of their affinity for promoter elements containing this sequence. We previously identified cDNA sequences representing four Arabidopsis thaliana type-TV zinc-finger proteins. Here we define the organisation and expression of GATA-1, GATA-2, GATA-3 and GATA-4 as well as DNA-binding characteristics of their encoded proteins. Transcripts from all four genes can be detected in all tissues examined suggesting that they are not developmentally regulated at the level of transcription. In vitro binding experiments with Escherichia coli-derived recombinant proteins were performed using motifs previously defined as targets for nuclear GATA-binding proteins. These studies reveal differences in DNA binding specificity of GATA-1 as compared to the other three proteins. In vivo protein-DNA interactions monitored by yeast one-hybrid assays reveal different binding characteristics as compared to those defined with E. coli-derived recombinant protein. Trans-activation of gene expression by the four Arabidopsis proteins via some, but not all, DNA elements tested indicates that the Arabidopsis proteins can form functional interactions with previously defined promoter elements containing GATA motifs. We conclude that the Arabidopsis type-IV zinc-finger proteins may represent the previously defined family of nuclear GATA-binding proteins implicated in light-responsive transcription.
许多光响应启动子含有GATA基序,并且已经鉴定出一些与这些元件相互作用的核蛋白。IV型锌指蛋白在动物和真菌中已得到广泛表征,由于它们对含有该序列的启动子元件具有亲和力,因此被称为GATA因子。我们之前鉴定了代表四种拟南芥IV型锌指蛋白的cDNA序列。在此,我们确定了GATA-1、GATA-2、GATA-3和GATA-4的结构与表达,以及它们编码蛋白的DNA结合特性。在所有检测的组织中都能检测到这四个基因的转录本,这表明它们在转录水平上不受发育调控。使用先前定义为核GATA结合蛋白靶标的基序,对源自大肠杆菌的重组蛋白进行了体外结合实验。这些研究揭示了GATA-1与其他三种蛋白在DNA结合特异性上的差异。通过酵母单杂交试验监测的体内蛋白质-DNA相互作用显示,与用源自大肠杆菌的重组蛋白所确定的结合特性不同。这四种拟南芥蛋白通过部分(而非全部)测试的DNA元件对基因表达进行反式激活,表明拟南芥蛋白可以与先前定义的含有GATA基序的启动子元件形成功能性相互作用。我们得出结论,拟南芥IV型锌指蛋白可能代表了先前定义的参与光响应转录的核GATA结合蛋白家族。