Quantification of prolyl cis-trans molecular switch as a timing device in auxin-regulated lateral root development in rice

脯氨酰顺反分子开关的定量作为生长素调节水稻侧根发育的定时装置

• 批准号: 1615350
• 负责人: Linda Nicholson
• 金额: $ 72.32万
• 依托单位: Cornell University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1615350&HistoricalAwards=false
• 关键词: Quantification; prolyl; cis; trans; molecular

This project explores a key part of the amazing molecular circuitry that governs how a seed develops into a whole plant, and addresses a major gap in our understanding of how dynamic motions in the molecular world can influence the shape of a living organism. In rice, a specific signal-responsive molecular circuit has been identified that controls the formation of lateral roots that branch off of the main vertical root. This research project will investigate relationships between the sprouting of lateral roots and the rate of a specific molecular switch that regulates the timing of this circuit, thereby linking this molecular rate to the formation of the mature plant. These investigations will foster training of graduate and undergraduate students across multidisciplinary fields, and will be incorporated into a graduate level course on protein structure and function. Importantly, a series of four learning modules that incorporate this scientific research will be produced and then disseminated via the vast network of 4-H chapters across New York state that reaches approximately 189,000 youth in regions that include low-income rural and American Indian populations. The primary potential societal impacts are to train young scientists in an exciting area of investigation, to contribute key insights into how molecular motions govern the development of whole organisms, and to stimulate interest in scientific exploration in youth who otherwise might not be exposed to concepts that span from molecules to plants. The overall goal of this project is to quantitatively investigate the role of a specific prolyl cis-trans molecular switch that acts as a timing device in an auxin-responsive transcription regulatory circuit that governs lateral root development in rice. The specific goals are to tune the molecular switching rate using gene editing, to measure induced changes in rate using NMR spectroscopy, to quantify the resulting changes in circuit dynamics in single cells using confocal fluorescence microscopy, and to observe corresponding changes in phenotype in the whole organism. The results of these experiments will be used to establish a quantitative mathematical model for prediction of the effects of this cis-trans switching rate on cellular dynamics, with impact on phenotype. The potentially transformative aspect of the project is that if successful, it will integrate knowledge across the scales from motions of individual bonds, to the dynamics of a molecular circuit that regulates gene transcription in a single cell, to a well-defined phenotype (i.e., "genotype-to-phenotype"). This project is jointly funded by the Molecular Biophysics and Systems and Synthetic Biology Clusters in the Division of Molecular and Cellular Biosciences.

该项目探索了令人惊叹的分子电路的关键部分，该电路控制种子如何发育成整个植物，并解决了我们对分子世界中的动态运动如何影响生物体形状的理解中的一个主要空白。 在水稻中，已经确定了一个特定的信号响应分子电路，它控制着从主垂直根分支出来的侧根的形成。该研究项目将调查侧根发芽与调节该电路时序的特定分子开关速率之间的关系，从而将该分子速率与成熟植物的形成联系起来。这些调查将促进跨多学科领域的研究生和本科生的培训，并将纳入蛋白质结构和功能的研究生课程。 重要的是，将制作一系列包含这一科学研究的四个学习模块，然后通过遍布纽约州的4-H分会的庞大网络传播，该网络覆盖包括低收入农村和美洲印第安人在内的地区的约189 000名青年。主要的潜在社会影响是在一个令人兴奋的研究领域培养年轻科学家，为分子运动如何控制整个生物体的发展提供关键见解，并激发年轻人对科学探索的兴趣，否则他们可能不会接触到从分子到植物的概念。本项目的总体目标是定量研究一个特定的脯氨酰顺反分子开关的作用，作为一个定时装置在生长素响应转录调控电路，管理水稻侧根发育。具体目标是使用基因编辑来调整分子转换速率，使用NMR光谱测量诱导的速率变化，使用共聚焦荧光显微镜量化单细胞中回路动力学的变化，并观察整个生物体中表型的相应变化。这些实验的结果将被用来建立一个定量的数学模型，预测这种顺式-反式转换率对细胞动力学的影响，对表型的影响。该项目的潜在变革方面是，如果成功，它将整合从单个键的运动到调节单细胞中基因转录的分子电路的动态，再到明确定义的表型（即，“基因型到表型”）。该项目由分子和细胞生物科学部的分子生物物理学和系统与合成生物学集群联合资助。