PHYTOCHROME--BIOGENESIS AND MOLECULAR PROPERTIES

植物色素——生物发生和分子特性

• 批准号: 3306978
• 负责人: Peter H. Quail
• 金额: $ 8.38万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-30 至 1996-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3306978
• 关键词: Arabidopsis; X ray crystallography; biological signal transduction; enzyme activity; gene deletion mutation; gene expression; genetic promoter element; genetic regulatory element; genetic transcription; genetically modified plants; immunocytochemistry; microinjections; monoclonal antibody; nonvisual photoreceptor; nucleic acid sequence; photobiology; plant genetics; protein kinase; site directed mutagenesis

Phytochrome is a regulatory master switch that controls gene expression in response to fight signals from the environment. The long term goal of the proposed research is to define the molecular mechanism by which this regulation is exerted. The specific objectives of this proposal are: (a) to identify sequence and structural features of the phytochrome A molecule responsible for its regulatory action; and (b) to define regulatory elements in the multidomain phyA gene promoter Arabidopsis responsible for differential expression from multiple transcription start sites. The experimental approaches will include: (a) biochemical analysis to assess the possibility that Avena phytochrome A has protein kinase activity similar to prokaryotic sensor proteins; (b) overexpression of mutagenized Avena phytochrome A in transgenic Arabidopsis and microorganisms to identify structurally and functionally important domains and residues; (c) microinjection of mutagenized phytochrome A into phytochrome-deficient transgenic Arabidopsis cells carrying phytochrome-responsive promoter-reporter fusion constructs to identify functionally critical sequences in the photoreceptor by transient expression assay; (d) production of homogeneous phytochrome A preparations by overexpression in microorganisms for X-ray crystallographic analysis; and (e) deletion and sequence substitution analysis of the Arabidopsis phyA promoter in transgenic Arabidopsis to delineate cis-acting DNA sequences involved in the regulation of this complex promoter. The significance of this research is that it will help define the mechanism by which a central regulatory molecule controls gene expression. Because phytochrome appears as yet to have no molecular counterpart in other eukaryotes, elucidation of this mechanism has the potential to broaden our understanding of the strategies used by eukaryotic cells to regulate gene expression in response to environmental stimuli.

光敏色素是控制基因表达的调节主开关 以响应来自环境的战斗信号。 的长期目标 拟议的研究是确定分子机制， 监管正在实施。 这项建议的具体目标是： 鉴定光敏色素A的序列和结构特征 负责其调节作用的分子;和（B）定义 拟南芥多结构域phyA基因启动子调控元件 负责从多个转录起始的差异表达 网站. 实验方法将包括：（a）生物化学分析， 评估燕麦光敏色素A具有蛋白激酶的可能性 活性类似于原核传感器蛋白;（B）过表达 在转基因拟南芥中诱变的燕麦光敏色素A， 微生物鉴定结构和功能重要 结构域和残留物;（c）显微注射诱变光敏色素A 转化到缺乏光敏色素的转基因拟南芥细胞中， 光敏色素响应启动子-报告基因融合构建体，以鉴定 功能关键序列在感光细胞的瞬时 （d）均质光敏色素A的产生 X射线用微生物过表达制剂 晶体学分析;和（e）缺失和序列取代 分析转基因拟南芥中的拟南芥phyA启动子， 描述参与调控这一过程的顺式作用DNA序列， 复合启动子 这项研究的意义在于，它将有助于定义 中心调节分子控制基因的机制 表情 因为光敏色素似乎还没有分子 在其他真核生物中，这种机制的阐明具有 有可能扩大我们对 真核细胞调节基因表达以响应环境 刺激。