Transcriptional Regulation of the Maize Anthocyanin Pathway

对玉米花青素途径的广泛遗传分析揭示了多个靶基因，这些基因的表达受多个调控基因的协调控制。我们的目标是了解编码花青素生物合成酶的基因转录受三个调控基因b、c1和a3协调控制的机制。拟议的体内和体外实验应该使我们能够开始区分调控蛋白如何相互作用以控制转录的可能模型。我们的具体目标是：1)突变地解剖它们相互作用所需的C1和B区域，2)识别涉及的其他蛋白质，3)建立分析B和C1蛋白/蛋白质相互作用的体外分析方法，4)评估B和C1蛋白是否直接与结构基因调控位点相互作用，5)确定B和C1调控未知结构基因启动子所需的顺式作用位点。6)使用可用的转座元件插入确定B的功能重要区域，7)确定A3是否在转录水平上起作用，如果是，克隆并鉴定A3。从这些研究中学到的基因调控原理应该适用于基因和分子相结合的方法尚不可行的其他系统。

Extensive genetic analyses of the maize anthocyanin pathway have revealed multiple target genes, whose expression is coordinately controlled by several regulatory genes. Our goal is to understand the mechanisms by which the transcription of the genes encoding the anthocyanin biosynthetic enzymes is coordinately controlled by three regulatory genes, b, c1, and a3. The proposed in vivo and in vitro experiments should enable us to begin to distinguish between possible models for how the regulatory proteins are interacting to control transcription. Our specific aims are to : 1) mutationally dissect the regions of C1 and B required for their interaction, 2) identify other proteins involved, 3) develop in vitro assays for analyzing B and C1 protein/protein interactions, 4) assess whether the B and C1 proteins interact directly with structural gene regulatory sites, 5) identify the cis-acting sites required for B and C1 regulation of uncharacterized structural gene promoters, 6) identify functionally important regions of B using available transposable element insertions, 7) determine if a3 is acting at the transcriptional level and if so, clone and characterize a3. %%% Principles of gene regulation learned from these studies should apply to other systems where a combined genetic and molecular approach is not yet feasible.