title: Expression of a bacterial bi-functional chorismate mutase/prephenate dehydratase modulates primary and secondary metabolism associated with aromatic amino acids in Arabidopsis.
abstract: Plants can synthesize the aromatic amino acid Phe via arogenate, but it is still not known whether they also use an alternative route for Phe biosynthesis via phenylpyruvate, like many micro-organisms. To examine this possibility, we expressed a bacterial bi-functional PheA (chorismate mutase/prephenate dehydratase) gene in Arabidopsis thaliana that converts chorismate via prephenate into phenylpyruvate. The PheA-expressing plants showed a large increase in the level of Phe, implying that they can convert phenylpyruvate into Phe. In addition, PheA expression rendered the plants more sensitive than wild-type plants to the Trp biosynthesis inhibitor 5-methyl-Trp, implying that Phe biosynthesis competes with Trp biosynthesis from their common precursor chorismate. Surprisingly, GC-MS, LC-MS and microarray analyses showed that this increase in Phe accumulation only had a very minor effect on the levels of other primary metabolites as well as on the transcriptome profile, implying little regulatory cross-interaction between the aromatic amino acid biosynthesis network and the bulk of the Arabidopsis transcriptome and primary metabolism. However, the levels of a number of secondary metabolites derived from all three aromatic amino acids (Phe, Trp and Tyr) were altered in the PheA plants, implying regulatory cross-interactions between the flux of aromatic amino acid biosynthesis from chorismate and their further metabolism into various secondary metabolites. Taken together, our results provide insights into the regulatory mechanisms of aromatic amino acid biosynthesis and their interaction with central primary metabolism, as well as the regulatory interface between primary and secondary metabolism.
authors.affiliation: Department of Plant Sciences, The Weizmann Institute of Science, Rehovot 76100, Israel.
authors.forename: Vered
authors.initials: V
authors.lastname: Tzin
authors.forename: Sergey
authors.initials: S
authors.lastname: Malitsky
authors.forename: Asaph
authors.initials: A
authors.lastname: Aharoni
authors.forename: Gad
authors.initials: G
authors.lastname: Galili
mesh_terms: D024322:Amino Acids, Aromatic / Q000378:metabolism*; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D002827:Chorismic Acid / Q000378:metabolism; D019295:Computational Biology; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D055432:Metabolomics; D009097:Multienzyme Complexes / Q000235:genetics / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D010649:Phenylalanine / Q000096:biosynthesis; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D011302:Prephenate Dehydratase / Q000235:genetics* / Q000378:metabolism; D018749:RNA, Plant / Q000235:genetics
chemical_list: D024322:Amino Acids, Aromatic; D029968:Escherichia coli Proteins; D009097:Multienzyme Complexes; C071216:P-protein, E coli; D018749:RNA, Plant; D010649:Phenylalanine; D011302:Prephenate Dehydratase; D002827:Chorismic Acid