A MAIZE PROTEIN ASSOCIATED WITH THE G-BOX BINDING COMPLEX HAS HOMOLOGY TO BRAIN REGULATORY PROTEINS
A MAIZE PROTEIN ASSOCIATED WITH THE G-BOX BINDING COMPLEX HAS HOMOLOGY TO BRAIN REGULATORY PROTEINS
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DOI：
10.1105/tpc.4.10.1295
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发表时间：
1992-10-01
期刊：
影响因子：
11.6
通讯作者：
FERL, RJ
中科院分区：
文献类型：
--
作者：
DEVETTEN, NC;LU, GH;FERL, RJ
G-box元件是许多诱导基因启动子的适度保守的组成部分，包括拟南芥和玉米的乙醇脱氢酶基因。我们使用针对部分纯化的 G 盒结合因子 (GBF) 活性生成的单克隆抗体来表征作为 DNA 结合复合物一部分的玉米蛋白。抗体与部分纯化的玉米 GBF 复合物相互作用，在凝胶阻滞测定中产生迁移较慢的复合物。免疫沉淀实验表明，抗体识别的蛋白质本身并不是 DNA 结合蛋白，而是与 DNA 结合复合物相关。这些单克隆抗体用于分离编码我们指定为 GF14 的蛋白质的 cDNA 克隆。玉米GF14含有类似亮氨酸拉链的区域以及酸性羧基和氨基末端，其中后者可以形成类似于已知的转录激活剂如VP16和GAL4的两亲性α螺旋。细胞培养物提取物的蛋白质凝胶印迹分析表明，大约 30 kD 的单一主要蛋白质被抗 GF14 识别；该蛋白质也主要存在于籽粒和根中。玉米 GF14 的推导氨基酸序列与拟南芥 GF14 和月见草 PHP-O 的同一性超过 80%，与一类哺乳动物脑蛋白（被描述为蛋白激酶 C 抑制剂以及酪氨酸和色氨酸羟化酶激活剂）的同一性超过 60%。 GF14 存在于多种单子叶植物和双子叶植物、裸子植物和酵母中。这表明与许多基因启动子区域中发现的核心序列相关的潜在调节蛋白的深层进化保守性。
The G-box element is a moderately conserved component of the promoter of many inducible genes, including the alcohol dehydrogenase genes of Arabidopsis and maize. We used monoclonal antibodies generated against partially purified G-box binding factor (GBF) activity to characterize maize proteins that are part of the DNA binding complex. Antibodies interacted with partially purified maize GBF complexes to produce a slower migrating complex in the gel retardation assay. Immunoprecipitation experiments suggested that the protein recognized by the antibody is not a DNA binding protein in and of itself, but rather is associated with the DNA binding complex. These monoclonal antibodies were used to isolate cDNA clones encoding a protein that we have designated GF14. Maize GF14 contains a region resembling a leucine zipper and acidic carboxy and amino termini, of which the latter can form an amphipathic alpha-helix similar to known transcriptional activators such as VP16 and GAL4. Protein gel blot analysis of cell culture extract showed that a single, major protein of approximately 30 kD is recognized by anti-GF14; the protein is also present predominantly in the kernel and root. The deduced amino acid sequence of maize GF14 is more than 80% identical to Arabidopsis GF14 and Oenothera PHP-O, and is more than 60% identical to a class of mammalian brain proteins described as both protein kinase C inhibitors and activators of tyrosine and tryptophan hydroxylases. GF14 is found in a variety of monocotyledons and dicotyledons, gymnosperms, and yeast. This suggests a deep evolutionary conservation of a potential regulatory protein associated with a core sequence found in the promoter region of many genes.