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

这个项目探索了令人惊叹的分子电路的一个关键部分，该电路支配着种子如何发育成一种完整的植物，并解决了我们在理解分子世界中的动态运动如何影响活的有机体的形状方面的一个主要空白。在水稻中，已经发现了一种特定的信号响应分子回路，它控制着从主垂直根分支而来的侧根的形成。这项研究项目将研究侧根萌发和特定分子开关速率之间的关系，该开关调节这一回路的时间，从而将该分子速率与成熟植物的形成联系起来。这些研究将促进跨多学科领域的研究生和本科生的培训，并将被纳入蛋白质结构和功能的研究生水平课程。重要的是，将制作一系列包含这项科学研究的四个学习单元，然后通过纽约州庞大的4-H分会网络进行传播，该网络覆盖了包括低收入农村人口和美国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训年轻科学家，就分子运动如何管理整个生物体的发育提供关键见解，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。这个项目的总体目标是定量研究特定的Pro顺反分子开关在控制水稻侧根发育的生长素响应转录调控电路中的计时装置的作用。具体目标是使用基因编辑来调整分子开关速率，使用核磁共振光谱来测量诱导的速率变化，使用共聚焦荧光显微镜来量化单个细胞中由此产生的电路动力学的变化，并观察整个生物体中相应的表型变化。这些实验的结果将被用来建立一个定量的数学模型，以预测这种顺式-反式转换率对细胞动力学的影响，以及对表型的影响。该项目潜在的变革性方面是，如果成功，它将整合从单个键的运动到调节单个细胞中基因转录的分子电路的动力学，再到定义明确的表型(即“基因到表型”)的各个层面的知识。该项目由分子和细胞生物科学部的分子生物学和系统与合成生物学集群共同资助。

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.