Novel auxin probes on auxin receptors and signal transduction

关于生长素受体和信号转导的新型生长素探针

基本信息

批准号：
18510197
负责人：
HAYASHI Ken-ichiro
金额：
$ 1.52万
依托单位：
Okayama University of Science
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

The regulation of gene expression by the hormone auxin is a crucial mechanism in plant development. Arabidopsis F-box protein TIR1 is a receptor for auxin and recent structural work has revealed the molecular mechanism of auxin perception. TIR1 is the substrate receptor of the ubiquitin-ligase complex SCF^<TIR1>. However, the detailed relations between signaling and physiological response are still unclear. We have developed two types of auxin probes to study auxin responses by chemical biological approach.1 : Novel synthetic auxin agonists and antagonists. These probes are specific to TIR1-mediated events in Arabidopsis and their mode of action in binding to TIR1 is confirmed by X-ray crystallographic analysis. Further we demonstrate the utility of these probes for the analysis of TIR1-mediated auxin signaling in the moss P. patens. Our work not only provides a new useful tool for plant chemical biology but also demonstrates the first example of a specific small molecule inhibitor of F-box protein-substrate recruitment.2: Novel auxin probes derived from natural products. Terfestatin A (TrfA), a novel auxin signaling inhibitor, was identified in a screen of Streptomyces sp. F40 extracts for inhibition of the expression of an auxin-inducible gene. However, the mode of action of TrfA has not been elucidated. To identify the active core structure, twenty-five derivatives of TrfA were synthesized and their inhibitory activities against auxin-inducible gene expression were evaluated. The structure-activity relationships revealed the essential active core structure of TrfA, 3-butoxy-4-methylbiphenyl-2, 6-diol, which will lead to the design of biotin-tagged active TrfA.

激素生长素对基因表达的调节是植物发育中的关键机制。拟南芥F-box蛋白TIR1是生长素的受体，最近的结构工作揭示了生长素感知的分子机制。 TIR1是泛素 - 连接酶复合物SCF^<TIR1>的底物受体。但是，信号传导和生理反应之间的详细关系尚不清楚。我们已经开发了两种类型的生长素探针来通过化学生物学方法研究生长素反应。1：新型的合成生长素激动剂和拮抗剂。这些探针特异性是拟南芥中TIR1介导的事件，其与TIR1结合的作用方式通过X射线晶体学分析证实。此外，我们证明了这些探针在Moss P. patens中分析TIR1介导的生长素信号传导的实用性。我们的工作不仅为植物化学生物学提供了一种新的有用工具，而且还展示了F-box蛋白质 - 基底募集的特定小分子抑制剂的第一个例子。2：来自天然产物的新型生长素探针。 Terfestatin A（TRFA）是一种新型的生长素信号抑制剂，在链霉菌链球菌的屏幕中鉴定出来。 F40提取物，用于抑制生长素诱导基因的表达。但是，尚未阐明TRFA的作用方式。为了识别活性核心结构，合成了TRFA的25个衍生物，并评估了其对生长素诱导基因表达的抑制活性。结构活性关系揭示了TRFA，3-丁克西-4-甲基苯基-2、6-二醇的基本活动核心结构，这将导致生物素标记的活性TRFA的设计。