DNA Recognition by Basic-Leucine Zipper Proteins

碱性亮氨酸拉链蛋白的 DNA 识别

• 批准号: 9206095
• 负责人: Mark Guiltinan
• 金额: $ 64.48万
• 依托单位: Pennsylvania State Univ University Park
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-08-15 至 1999-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9206095&HistoricalAwards=false
• 关键词: DNA; Recognition; Basic; Leucine; Zipper

EmBP-1 is a wheat basic-leucine zipper (bZIP) class transcription factor which has been implicated in the control of gene expression. Proposed experimentation will address the structure-function relationships of the DNA binding domain of EmBP-1. By analyzing mutant DNA binding proteins with altered binding preferences, insight can be gained of the ways in which amino acid sequences within the DNA binding domains of bZIP proteins encode functional specificities. The minimal domains for the dimerization and DNA binding functions of EmBP-1 will be determined using deletion analysis and in vitro binding assays. cDNA libraries encoding randomly mutagenized basic region domains will be constructed and probed with degenerate oligonucleotides to determine how changes in the amino acid sequences of the bZIP domains are manifested in altered binding specificities. The functional significance of these findings will be tested in vivo by expressing mutagenized versions of EmBP-1 in transgenic Arabidopsis plants. Additional, to examine the natural variation in bZIP sequences, members of the EmBP-1 gene family will be isolated from Arabidopsis. Because the leucine zipper class of transcription factors has been conserved from plants to animals, the knowledge gained from the proposed research will be applicable to the basic principles operating in most eukaryotic cells. %%% EmBP-1 is a wheat DNA binding protein involved in controlling gene expression. Understanding the way in which DNA binding proteins recognize specific DNA sequences will lead to a greater understanding of the mechanisms by which cells activate gene expression. This has great significance in understanding normal growth and development as well as in understanding aberrant gene expression which is often associated with human disease such as cancer. By analyzing mutant DNA binding proteins with altered binding preferences, insight can be gained of the ways in which this class of proteins function. Randomly mutagenizeds Dna binding domains will be constructed analyzed to determine how changes in the amino acid sequences are manifested in altered binding specificity. These findings will be tested by expressing mutagenized versions of EnBP-1 in genetically engineered plants, Additionally, to examine the natural variation in DNA binding sequences, related genes will be isolated from Arabidopsis (a model plant species used in molecular genetic studies). Because this type of protein is found within organisms ranging from plants to animals, the knowledge gained from the proposed research will be applicable to the basic principles operating in most cells.

EmBP-1是小麦碱性亮氨酸拉链（bZIP）类 转录因子，该转录因子与控制 的基因表达。 拟议的实验将解决 的DNA结合结构域的结构-功能关系 EmBP-1。 通过分析突变的DNA结合蛋白， 约束偏好，可以获得洞察的方式， bZIP的DNA结合结构域内的氨基酸序列 蛋白质编码功能特异性。 极小域 对于EmBP-1的二聚化和DNA结合功能， 使用缺失分析和体外结合来确定 测定。 编码随机诱变的碱性磷酸酶的cDNA文库 区域域将被构建并用简并探针探测。 寡核苷酸来确定氨基酸的变化 bZIP结构域的序列表现为改变的结合 特殊性 这些发现的功能意义 将通过表达突变形式的 转基因拟南芥植物中的EmBP-1。 此外， 研究bZIP序列的自然变异， EmBP-1基因家族将从拟南芥中分离。 因为 亮氨酸拉链类转录因子已经被 从植物到动物，从植物中获得的知识 建议研究将适用的基本原则 在大多数真核细胞中运作。 %%% EmBP-1是一种小麦DNA结合蛋白， 基因表达。 了解DNA结合的方式 蛋白质识别特定的DNA序列， 更好地理解细胞激活的机制 基因表达。 这对于理解 正常的生长和发育以及理解 异常基因表达通常与人类 疾病，如癌症。 通过分析突变DNA结合 蛋白质与改变结合偏好，洞察力可以 获得了这类蛋白质的功能方式。 将构建随机诱变的DNA结合结构域 分析以确定氨基酸序列的变化 表现为结合特异性的改变。 这些发现 将通过在大肠杆菌中表达诱变形式的EnBP-1来测试。 基因工程植物，此外，检查 DNA结合序列的自然变异，相关基因将 分离自拟南芥（Arabidopsis）（一种用于 分子遗传学研究）。 因为这种蛋白质是 在从植物到动物的生物体中发现， 从拟议的研究中获得的知识将是适用的 大多数细胞的基本原理。