ABRB BINDING TO DEVELOPMENTALLY CONTROLLED PROMOTERS

首席研究员建议研究芽孢杆菌的abr B蛋白 枯草杆菌。AbrB是调节基因表达的一个关键的全球调节因子 为了适应次优环境中的新陈代谢需求，压力和最初 产孢子发育过程的各个阶段。除了……之外 防止相关固定相的不适当表达 在快速增长期间发挥作用，最近的证据表明，abr B 也在调节分解代谢抑制方面发挥作用，并可能影响 依赖于增长率的翻译成分的调节 仪器。AbrB是一种转录调节因子，其N-末端结构域 代表了一种新的DNA结合基序，它主要识别微妙的 由一个子集假设的三维DNA结构 不同的序列。国际刑事法院辩称，对这些因素的解释 负责灵活的AbrB结合专一性将在 蛋白质-DNA识别机制和细胞如何经济地 使用一种蛋白质来协调各种应激反应和 发展选择。目前，超过35个基因编码一个广泛的阵列 代谢功能，包括一些正常产孢子所必需的， 已知有AbrB结合位点，通常在启动子区域。 为了更好地定义DNA靶标的确切结构参数 被AbrB识别，与选定位点的结合将被HIGH探测 分辨率足迹技术和动力学研究使用 灵敏的实时检测。蛋白质结构与DNA的关系 结合突变分析将探测结合属性， 分离的DNA结合区的核磁共振，C区多聚体的研究 和AbrB-DNA复合体的结晶学研究。体外转录和 结合分析将检验有关AbrB介导的假说 代表性发起人的监管机制不同之处 AbrB依赖效应的类别是可疑的。如有需要， 将获得体内确凿的遗传学证据。

The principal investigator proposes to study the AbrB protein Bacillus subtilis. AbrB is a key global regulator which adjusts gene expression to fit metabolic needs in suboptimal environments, stress and the initial stages of the developmental process of sporulation. In addition to preventing inappropriate expression of stationary phase associated functions during rapid growth, recent evidence suggests that AbrB also plays a role in modulating catabolite repression and could affect growth-rate dependent regulation of components of the translational apparatus. AbrB is a transcriptional regulator whose N-terminal domain represents a novel DNA-binding motif that primarily recognizes subtle three-dimensional DNA structures that are assumed by a subset of varying sequences. The PI argues that elucidation of the factors responsible for flexible AbrB binding specificity will provide insights in protein-DNA recognition mechanisms and how a cell can economically use a single protein to coordinate a variety of stress responses and developmental options. Currently, over 35 genes encoding a wide array of metabolic functions, including some essential for normal sporulation, are known to have AbrB binding sites, usually in the promoter regions. To better define the exact structural parameters of the DNA targets recognized by AbrB, binding to select sites will be probed by high resolution footprinting techniques and kinetics examined using a sensitive real-time assay. The relationship of protein structure to DNA- binding properties will be probed by a combination o mutant analysis, NMR of isolated DNA-binding domain, C domain multimerization studies and crystallography of AbrB-DNA complexes. In vitro transcription and binding assays will test hypotheses regarding AbrB-mediated regulatory mechanisms at representative promoters where different classes of AbrB-dependent effects are suspected. Where necessary, confirmatory in vivo genetic evidence will be obtained.