Arabidopsis 2010: High-Throughput Analysis of the Mystery Genes of CoA Metabolism

这个2010年的项目将确定59个辅酶a代谢基因的具体功能。其中编码羧基辅酶a连接酶的基因44个，编码羧基辅酶a硫酯酶的基因15个。目前，这59个基因中只有11个已知功能。辅酶a连接的底物是中间代谢和产生细胞及其细胞器的结构构建块的途径的核心，许多保护植物免受生物和非生物胁迫的化学物质，以及几种激素。循环碳的分解代谢途径也依赖于辅酶a。许多辅酶a途径对生化和遗传学研究具有抗性。这个项目很重要，因为它将为基因共同贡献的许多途径提供生化和遗传接入点。确定基因的明确功能将需要并行/集成部署四种策略：1)生物信息学方法，包括转录物分析，启动子基序分析和蛋白质结构建模。2)重组蛋白的生化分析将确定底物并确定酶动力学。3)组织特异性表达和亚细胞靶向信息将区分利用不同同工酶的途径。4)反向遗传学（基因敲除/RNAi/TILLING）和过表达将定义更广泛的生物学背景。这些结果将为了解拟南芥的一些生化途径和其他途径提供重要的初步信息。更广泛的影响：由于许多与辅酶a相关的途径目前尚不清楚，该项目的结果将促进植物生物学不同领域的研究。在这个项目下开发的资料将直接提供（http://www.ibc.wsu.edu/research/browse/NSF2010），并纳入AraCyc和其他TAIR数据库。生物资源将通过库存中心提供。最终，更好地了解59个基因是这项研究的重点，将有助于提高农业、林业和环境管理许多领域的能力。与工业界的密切接触将有助于这些更广泛的影响。华盛顿州立大学对学生培训的多样性有着坚定的承诺，并有几个积极的项目招收少数民族和代表性不足的群体。研究生和本科生都将通过参与这项研究项目，获得研究技术和科学逻辑方面的培训。

This 2010 project will determine specific functions for 59 genes of CoA metabolism. The genes include 44 encoding carboxy-CoA ligases and 15 encoding carboxy-CoA thioesterases. Currently, only 11 of these 59 genes have known functions. CoA-linked substrates are central to intermediary metabolism and to pathways that produce structural building blocks of the cell and its organelles, many chemicals that protect plants against biotic and abiotic stresses, and several hormones. Catabolic pathways that recycle carbon also depend upon CoA. Many CoA pathways have been resistant to biochemical and genetic investigation. This project is important because it will provide biochemical and genetic access points in many pathways to which the genes collectively contribute. Assigning definitive functions for the genes will require parallel/integrated deployment of four strategies: 1) Bioinformatics approaches including transcript profiling, promoter motif analysis, and modeling of protein structures. 2) Biochemical assays of recombinant proteins will identify substrates and define enzyme kinetics. 3) Tissue-specific expression and subcellular-targeting information will distinguish pathways utilizing different isozymes. 4) Reverse genetics (knockout/RNAi/TILLING as appropriate) and overexpression will define the broader biological context. These results will provide critical first openings into a number of biochemical pathways in Arabidopsis and new information about others.Broader Impacts: Because many CoA-linked pathways are poorly understood currently, the results from this project will facilitate investigations in diverse areas of plant biology. Information developed under this project will be available directly (http://www.ibc.wsu.edu/research/browse/NSF2010) and incorporated into AraCyc and other TAIR databases. Biological resources will be available through the Stock Centers. Eventually, better understanding of the 59 genes that are the focus of this research will contribute to improved capabilities in many areas of agriculture, forestry and environmental management. These broader impacts will be assisted by close contacts with industry. Washington State University has a strong commitment to diversity in student training, and has several active programs for recruitment of minority and underrepresented groups. Both graduate and undergraduate students will receive training in research techniques and scientific logic through their involvement in this research project.