Transcriptional Regulation by the P Gene, a Myb-Homolog in Maize

在以前的工作中，我们已经证明了P基因从类黄酮生物合成途径的三个基因调控RNA的积累。我们还表明，P编码的蛋白质与癌基因蛋白c-myb家族的DNA结合域具有显著的同源性。在这里，我们提出结合分子，生化和遗传分析的P基因产物的调控功能。我们将产生针对P蛋白的抗体，并利用这些抗体来鉴定P编码蛋白，并确定翻译后修饰可能参与P的调节功能，以及P编码表达在不同玉米组织中P转录本的作用。磷调控基因中所必需的靶序列将通过功能测试和体外DNA结合测定来确定。最后，我们将使用两个互补的体内系统分析p编码蛋白的突变。首先，我们将尝试在酿酒酵母中复制P对目标基因的调控；如果成功,这种方法将有助于快速分析磷调控，并允许鉴定与磷相互作用的转录因子。第二个体内系统利用转座元件Ac在磷蛋白的特定位点产生突变等位基因来检测磷在玉米植物中的功能。​总之，这些结果将有助于揭示植物调控基因激活转录的基本机制

In previous work we have shown that the P gene regulates the accumulation of RNA from three genes of the flavonoid biosynthetic pathway. We have also shown that P encodes proteins with significant homology to the DNA binding domain of the c-myb family of oncogene proteins. Here, we propose a combined molecular, biochemical and genetic analysis of the regulatory function of the P gene products. We will generate antibodies against P proteins and use these to identify the P-encoded proteins and determine post-translational modifications may be involved in the regulatory function of P. The role of the P-encoded expression of the P transcripts in different maize tissues. The target sequences present in P regulated genes necessary for P regulation will be determined by functional testing and in vitro DNA binding assays. Finally, we will analyze mutations in the P-encoded proteins using two complementary in vivo systems. In the first, we will attempt to reproduce P regulation of a target gene in the yeast S. cerevisiae; if successful, this approach will facilitate rapid analysis of P regulation and allow the identification of transcription factors which interact with P. The second in vivo system examines P function in the maize plant using the transposable element Ac to generate mutant alleles at the specific sites in the P protein. Together, these results should help to reveal fundamental mechanisms by which a plant regulatory gene activates transcription.***//