G Protein Activation through Uncoupling Regulator of G Signaling Protein, AtRGS1

智力优势：为了使生物体正常发育并对其环境作出适当的反应，组成生物体的细胞必须相互交流。细胞用来交流的信号可以是激素、光、小分子、电脉冲和触觉。例如，光可以打开（激活）一个光感受器，然后激活一些其他的分子复合物，然后激活更多的开关，以此来给细胞带来变化。在受体的信号和细胞中发生的变化之间是一个叫做异源三聚体G蛋白复合物的分子开关。异三聚体G蛋白偶联信号用于生物体的正常发育以及对引起疾病的病原体的反应。最近对异三聚体G蛋白偶联信号传导的研究表明，信号激活如何调节的潜在范式转变。例如，在后生动物和真菌中，激活是限速步骤，由配体刺激的细胞表面G蛋白偶联受体（GPCR）催化。相比之下，植物调节GTP水解导致静息状态的反向反应，而在拟南芥中，这种静息状态是由一种名为AtRGS1的G信号（RGS）跨膜调节蛋白维持的。在拟南芥中，d -葡萄糖和/或糖代谢产物是部分由异源三聚体G蛋白复合物介导的信号。假设葡萄糖和/或糖代谢产物通过调节AtRGS1和AtGPA1之间的偶联来激活G蛋白途径，并且已经证明d -葡萄糖诱导AtRGS1的快速内化，而不是AtGPA1。要回答的关键问题包括如何实现解偶联、解偶联的结果以及解偶联的分子机制。这些问题将使用数学模型，信号组件的遗传操作和显微镜检查。更广泛的影响：更广泛的影响是高中生物学学生对如何通过信号转导实现差异细胞结果的教育。与北卡罗来纳大学外展计划合作，将进一步开发一个名为“相同基因-不同命运”的教学模块，以教授差异基因表达的概念以及具有相同基因的一组细胞的不同细胞命运的基础。虽然该模块目前是为生物I和II学生（9年级和10年级）设计的，但它适用于AP生物和大学新生。这个项目将通过在指令中加入表观遗传学和信号诱导变化等概念，使这些适应适应更高的主题。该模块的湿实验室部分利用转基因植物的基因启动子驱动酶活性，使植物的不同部分变成蓝色。蓝色区域取决于基因启动子的类型，并说明不同的基因表达。将举办相关的教师研讨会，以教授和推广该模块的使用。该模块将首先在北卡罗来纳州的高中分发，但最终将在全国范围内分发。

Intellectual Merit: For organisms to develop normally and to react appropriately to their environment, the cells that compose organisms must communicate with each other. Signals used by cells to communicate can be hormones, light, small molecules, electrical impulses, and touch. For example, light can turn on (activate) a light receptor which then activates a few other molecular complexes that then activates many more switches, and so on to afford changes in the cell. Between the signal at the receptor and the changes occurring in the cell is a molecular switch called the heterotrimeric G protein complex. Heterotrimeric G protein-coupled signaling is used for normal development of the organism as well as reactions to pathogens causing disease. Recent investigations of heterotrimeric G protein-coupled signaling using divergent model systems reveals a potential paradigm shift on how signal activation can be regulated. For example, in metazoans and fungi, activation is the rate limiting step and is catalyzed by a ligand-stimulated, cell surface G protein coupled receptor (GPCR). In contrast, plants regulate the back reaction which is GTP hydrolysis leading to the resting state, and in Arabidopsis, this resting state is maintained by a transmembrane regulator of G Signaling (RGS) protein designated AtRGS1. In Arabidopsis, D-glucose and/or sugar metabolites are signals that are mediated in part by the heterotrimeric G protein complex. It is hypothesized that glucose and/or sugar metabolites activate the G protein pathway by regulating the coupling between AtRGS1 and AtGPA1, and it has been demonstrated that D-glucose induces a rapid internalization of AtRGS1, but not AtGPA1. Key questions to be answered include how uncoupling is achieved, the consequence of uncoupling, and the molecular mechanism for uncoupling. These questions will be examined using mathematical modeling, genetic manipulation of the signaling components, and microscopy. Broader Impacts: The broader impact is education of high school biology students on how differential cellular outcomes can be achieved through signal transduction. In collaboration with the UNC outreach program, a teaching module called "Same Genes- Different Fates" will be further developed to teach the concept of differential gene expression and the basis for different cell fates for a set of cells all having the same genes. While the module is currently designed for Biology I and II students (9th and 10th graders), it is adaptable to AP biology and college freshmen. This project will make those adaptations to the higher subject matter by adding concepts like epigenetics and signaling-induced changes into the instructions. The wet-lab part of the module utilizes transgenic plants that have gene promoters driving an enzyme activity that causes different parts of the plant to turn blue. The area of blue is dependent on the type of gene promoter and illustrates differential gene expression. Associated teacher workshops will be held to teach and promote the use of this module. The module will be distributed to high schools initially in North Carolina but eventually nationwide.