title: Arabidopsis and maize RidA proteins preempt reactive enamine/imine damage to branched-chain amino acid biosynthesis in plastids.
abstract: RidA (for Reactive Intermediate Deaminase A) proteins are ubiquitous, yet their function in eukaryotes is unclear. It is known that deleting Salmonella enterica ridA causes Ser sensitivity and that S. enterica RidA and its homologs from other organisms hydrolyze the enamine/imine intermediates that Thr dehydratase forms from Ser or Thr. In S. enterica, the Ser-derived enamine/imine inactivates a branched-chain aminotransferase; RidA prevents this damage. Arabidopsis thaliana and maize (Zea mays) have a RidA homolog that is predicted to be plastidial. Expression of either homolog complemented the Ser sensitivity of the S. enterica ridA mutant. The purified proteins hydrolyzed the enamines/imines formed by Thr dehydratase from Ser or Thr and protected the Arabidopsis plastidial branched-chain aminotransferase BCAT3 from inactivation by the Ser-derived enamine/imine. In vitro chloroplast import assays and in vivo localization of green fluorescent protein fusions showed that Arabidopsis RidA and Thr dehydratase are chloroplast targeted. Disrupting Arabidopsis RidA reduced root growth and raised the root and shoot levels of the branched-chain amino acid biosynthesis intermediate 2-oxobutanoate; Ser treatment exacerbated these effects in roots. Supplying Ile reversed the root growth defect. These results indicate that plastidial RidA proteins can preempt damage to BCAT3 and Ile biosynthesis by hydrolyzing the Ser-derived enamine/imine product of Thr dehydratase.
authors: Horticultural Sciences Department, University of Florida, Gainesville, Florida 32611 tomniehaus@ufl.edu.
authors: Thomas D
authors: TD
authors: Niehaus
authors: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario N1G 2W1, Canada.
authors: Thuy N D
authors: TN
authors: Nguyen
authors: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario N1G 2W1, Canada.
authors: Satinder K
authors: SK
authors: Gidda
authors: Metabolics Core, UC Davis Genome Center, University of California, Davis, California 95616.
authors: Mona
authors: M
authors: ElBadawi-Sidhu
authors: Department of Microbiology, University of Georgia, Athens, Georgia 30602.
authors: Jennifer A
authors: JA
authors: Lambrecht
authors: Horticultural Sciences Department, University of Florida, Gainesville, Florida 32611.
authors: Donald R
authors: DR
authors: McCarty
authors: Department of Microbiology, University of Georgia, Athens, Georgia 30602.
authors: Diana M
authors: DM
authors: Downs
authors: Department of Biochemistry and Molecular Biology, New York Medical College, Valhalla, New York 10595.
authors: Arthur J L
authors: AJ
authors: Cooper
authors: Metabolics Core, UC Davis Genome Center, University of California, Davis, California 95616.
authors: Oliver
authors: O
authors: Fiehn
authors: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario N1G 2W1, Canada.
authors: Robert T
authors: RT
authors: Mullen
authors: Horticultural Sciences Department, University of Florida, Gainesville, Florida 32611.
authors: Andrew D
authors: AD
authors: Hanson
mesh_terms: D000595:Amino Acid Sequence; D000597:Amino Acids, Branched-Chain / Q000378:metabolism*; D000619:Aminohydrolases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D017360:Arabidopsis / Q000737:chemistry / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D002087:Butyrates / Q000378:metabolism; D006868:Hydrolysis; D007097:Imines / Q000378:metabolism; D012695:L-Serine Dehydratase / Q000235:genetics / Q000378:metabolism*; D055432:Metabolomics; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D018517:Plant Roots / Q000737:chemistry / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D018520:Plant Shoots / Q000737:chemistry / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D018087:Plastids / Q000201:enzymology; D016415:Sequence Alignment; D013913:Threonine Dehydratase / Q000235:genetics / Q000378:metabolism*; D000637:Transaminases / Q000235:genetics / Q000378:metabolism*; D003313:Zea mays / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D000597:Amino Acids, Branched-Chain; D029681:Arabidopsis Proteins; D002087:Butyrates; D007097:Imines; D010940:Plant Proteins; C005087:alpha-ketobutyric acid; D000637:Transaminases; C530160:branched-chain aminotransferase-3, Arabidopsis; D000619:Aminohydrolases; C000599350:RidA protein, Arabidopsis; D012695:L-Serine Dehydratase; D013913:Threonine Dehydratase