The GRIK-SnRK1 Protein Kinase Cascade And Its Potential Role In Regulating TCP Transcription Factors In Arabidopsis

知识优势：tcp构成了一个大型的植物特异性转录因子家族，由保守的基本螺旋-环-螺旋结构域定义。它们分为两类，一类促进细胞增殖，二类促进叶片和花的分化。tcp是序列特异性的DNA结合蛋白，以同二聚体或异二聚体的形式与共享或重叠的基序结合。考虑到TCP家族的复杂性及其组合功能的潜力，它很可能受到一系列复杂的监管机制的管理。该项目的总体目标是检查GRIK-SnRK1蛋白激酶级联在TCP调节中的作用。尽管SnRK1以控制和协调植物的碳代谢和能量平衡而闻名，但越来越多的证据表明，它也在植物发育中发挥重要作用，并通过磷酸化转录因子控制植物基因表达。激活SnRK1的GRIK仅在植物幼嫩组织、培养细胞和双病毒感染细胞中积累，所有这些细胞都复制植物染色体DNA。拟南芥TCP的一个子集被GRIK-SnRK1激酶级联磷酸化，一类TCP结合基序在snrk1下调基因中富集。该项目将表征所选TCP的GRIK-SnRK1靶标残基，并检查改变其磷酸化状态对TCP功能的影响。这项研究是了解植物中tcp如何受磷酸化调控以及翻译后机制在调节这一独特转录因子家族中的作用的重要的第一步。更好地了解TCP调控机制和GRIK-SnRK1级联在TCP调控中的作用，可能会导致开发新的植物生长调控策略，特别是在代谢胁迫条件下。更广泛的影响：将分子、细胞和生化技术结合应用于TCP磷酸化，为本科生、研究生和高级研究人员提供了独特的培训机会。pi将共同努力，促进实验室之间的智力和协作互动，以确保参与者得到广泛的培训，并发展将综合方法应用于复杂生物学问题所需的技能。项目参与者还将与当地教育工作者和北卡罗来纳州生命与科学博物馆合作，为中学教师提供为期两天的“手提箱里的科学”基因工具包研讨会。该工具包被设计为辅助教学辅助教师着手在中学年级遗传学内容，并对准NC标准课程的研究，以确保最大限度的潜在使用。研讨会主题将包括工具包内容，NC标准课程的学习要求，课堂实施，中级水平的遗传科学概述，以及学习策略的评估。外展项目有可能覆盖北卡罗来纳州格兰维尔县和达勒姆县的3600名中学生，这两个县都有大量少数民族和经济上处于不利地位的学生。更广泛的影响包括聘请30名中学数学和科学教师参加讲习班，通过博物馆工作人员提供后续支持，并在下一学年向参与的教师免费提供工具包。

Intellectual Merit: The TCPs constitute a large family of plant-specific transcription factors that are defined by a conserved basic helix-loop-helix domain. They fall into two classes with Class I TCPs promoting cell proliferation, and Class II members promoting leaf and floral differentiation. TCPs are sequence-specific DNA binding proteins that bind to shared or overlapping motifs as homo- or heterodimers. Given the complexity of the TCP family and its potential to function combinatorially, it is likely to be governed by an intricate array of regulatory mechanisms. The overall goal of the project is to examine the role of the GRIK-SnRK1 protein kinase cascade in TCP regulation. Although SnRK1 is best known for controlling and coordinating carbon metabolism and energy balance in plants, there is increasing evidence that it also plays an essential role in plant development and controls plant gene expression by phosphorylating transcription factors. GRIK, which activates SnRK1, only accumulates in young plant tissues, cultured cells and geminivirus-infected cells, all of which replicate plant chromosomal DNA. A subset of Arabidopsis TCPs are phosphorylated by the GRIK-SnRK1 kinase cascade, and Class I TCP binding motifs are enriched in SnRK1-down regulated genes. The project will characterize GRIK-SnRK1 target residues of selected TCPs and examine the consequence of altering their phosphorylation status on TCP function. This research represents an important first step in understanding how TCPs are regulated by phosphorylation and the role of post-translational mechanisms in modulating this unique transcription factor family in plants. A better understanding of TCP regulatory mechanisms and the role of the GRIK-SnRK1 cascade in TCP control could lead to the development of new strategies for modulating plant growth, especially under metabolic stress conditions. Broader Impacts: The combined application of molecular, cellular and biochemical techniques to TCP phosphorylation provides a unique training opportunity for undergraduate, graduate and senior researchers. The PIs will work together to foster intellectual and collaborative interactions between their laboratories to ensure that participants are broadly trained and develop the skills necessary to apply integrated approaches to complex biological problems. Project participants will also work with local educators and the NC Museum of Life and Science to offer a two-day workshop based on a genetic "Science in a Suitcase" kit for middle school teachers. The kit is designed as a supplemental teaching aid for teachers embarking on genetics content in the middle school grades and is aligned to the NC Standard Course of Study to ensure maximum potential use. Workshop topics will include kit contents, the NC Standard Course of Study requirements, classroom implementation, overview of genetic science at the middle grades level, and assessment of learning strategies. The outreach project has the potential to reach 3,600 middle school students in NC Granville and Durham Counties, both of which have large numbers of minority and economically disadvantage students. The broader impacts include engaging 30 middle grades math and science teachers in the workshops, providing follow-up support through the Museum staff, and providing the kit at no-cost to participating teachers during the following academic year.