Peters D G, Caddick M X
Department of Genetics and Microbiology, Donnan Laboratories, University of Liverpool, UK.
Nucleic Acids Res. 1994 Dec 11;22(24):5164-72. doi: 10.1093/nar/22.24.5164.
In Aspergillus nidulans the regulatory gene areA is responsible for mediating nitrogen metabolite repression. The areA product (AREA) represents an example of the GATA family of DNA binding proteins, which are characterised by the presence of a GATA domain consisting of a zinc finger within a highly conserved region of 52 amino acids. Among the other transcription factors included in this family is the principal erythroid transcription factor, GATA-1, which contains two GATA domains. In order to demonstrate high specificity binding of native AREA to DNA containing the sequence -GATA-, and investigate the presence in A.nidulans of other proteins with related specificities, we have used gel mobility shift assays. Both AREA-dependent and independent complexes have been identified. Two strains bearing chimeric genes were also characterised. In these, the region encoding the native GATA domain of AREA was replaced by sequences from murine GATA-1 cDNA encoding either the equivalent C-terminal domain or both the N and C-terminal domains. Strains bearing the areA::NC-GATA construct, which includes the sequence encoding both the N and C-terminal domains of GATA-1, leads to a pronounced increase in one of two AREA-dependent complexes and implicates the N-terminal domain of GATA-1 in mediating protein-protein interactions.
在构巢曲霉中，调控基因areA负责介导氮代谢物阻遏。areA产物（AREA）是DNA结合蛋白GATA家族的一个例子，该家族的特征是存在一个由52个氨基酸的高度保守区域内的锌指组成的GATA结构域。该家族中包括的其他转录因子之一是主要的红细胞转录因子GATA-1，它含有两个GATA结构域。为了证明天然AREA与含有-GATA-序列的DNA的高特异性结合，并研究构巢曲霉中其他具有相关特异性的蛋白质的存在情况，我们使用了凝胶迁移率变动分析。已鉴定出依赖AREA和不依赖AREA的复合物。还对两个携带嵌合基因的菌株进行了表征。在这些菌株中，编码AREA天然GATA结构域的区域被来自小鼠GATA-1 cDNA的序列取代，该序列编码等效的C末端结构域或N和C末端结构域。携带areA::NC-GATA构建体的菌株，其中包括编码GATA-1的N和C末端结构域的序列，导致两个依赖AREA的复合物之一显著增加，并表明GATA-1的N末端结构域参与介导蛋白质-蛋白质相互作用。
