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

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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.