TRANS-ACTING FACTORS REGULATING ADH GENE EXPRESSION

本提案的总体目标是深入了解 DNA结构，反式作用因子，蛋白质， 核酸相互作用和基因调控 植物中的转录 实验涉及玉米 乙醇脱氢酶基因 Adh 1和Adh 2表现出差异 组织特异性，但厌氧介导的协调 两种Adh基因的转录表达。 这提供了 独特的机会，以评估所发挥的作用， 激活中的因子和蛋白质-核酸相互作用 两个密切相关的基因，并比较和对比 基因转录的发育和环境激活。 这一相当直接的提议的主要焦点是， 反式作用的体外表征和纯化 影响Adh基因调节的蛋白质因素。 虽然这一一般研究领域一直在积极进行， 动物系统，植物反式作用因子才刚刚开始 接受调查 是什么让这个提议特别 可行和适当的是，我们已经知道（至少一些 的）Adh调节中涉及的因素。 从我们的体内 足迹研究我们知道至少四个结合位点 调节因子，我们知道它们的硫酸二甲酯足迹， 签名. 因此，我们确切地知道我们是什么因素， 他们不只是寻找那些 与Adh DNA结合。

The overall goal of this proposal is to gain insight into the relationships among DNA structure, trans-acting factors, protein- nucleic acid interactions, and the regulation of gene transcription in plants. The experiments involve the maize alcohol dehydrogenase genes. Adhl and Adh2 exhibit differential tissue specificity, yet anaerobiosis mediates the coordinate transcriptional expression of both Adh genes. This provides a unique opportunity to evaluate the roles played by trans-acting factors and protein-nucleic acid interactions in the activation of two closely related genes, and to compare and contrast the developmental and environmental activation of gene transcription. The main focus of this rather straightforward proposal is the in vitro characterization and purification of the trans-acting protein factors that affect the regulation of the Adh genes. While this general area of research has been actively pursued in animal systems, plant trans-acting factors are only now beginning to be investigated. What makes this proposal particularly feasible and appropriate is that we already know (at least some of) the factors involved in Adh regulation. From our in vivo footprinting studies we know the binding sites of at least four regulatory factors, and we know their dimethyl sulfate footprint signatures. Therefore we know precisely the factors we are searching to purify and are not simply looking for proteins that (just) bind to Adh DNA.