title: root uv-b sensitive mutants are suppressed by specific mutations in ASPARTATE AMINOTRANSFERASE2 and by exogenous vitamin B6.
abstract: Vitamin B6 (vitB6) serves as an essential cofactor for more than 140 enzymes. Pyridoxal 5'-phosphate (PLP), active cofactor form of vitB6, can be photolytically destroyed by trace amounts of ultraviolet-B (UV-B). How sun-exposed organisms cope with PLP photosensitivity and modulate vitB6 homeostasis is currently unknown. We previously reported on two Arabidopsis mutants, rus1 and rus2, that are hypersensitive to trace amounts of UV-B light. We performed mutagenesis screens for second-site suppressors of the rus mutant phenotype and identified mutations in the ASPARTATE AMINOTRANSFERASE2 (ASP2) gene. ASP2 encodes for cytosolic aspartate aminotransferase (AAT), a PLP-dependent enzyme that plays a key role in carbon and nitrogen metabolism. Genetic analyses have shown that specific amino acid substitutions in ASP2 override the phenotypes of rus1 and rus2 single mutants as well as rus1 rus2 double mutant. These substitutions, all shown to reside at specific positions in the PLP-binding pocket, resulted in no PLP binding. Additional asp2 mutants that abolish AAT enzymatic activity, but which alter amino acids outside of the PLP-binding pocket, fail to suppress the rus phenotype. Furthermore, exogenously adding vitB6 in growth media can rescue both rus1 and rus2. Our data suggest that AAT plays a role in vitB6 homeostasis in Arabidopsis.
authors.affiliation: Department of Biology, 1600 Holloway Avenue, San Francisco State University, San Francisco, CA 94132, USA.
authors.forename: Colin D
authors.initials: CD
authors.lastname: Leasure
authors.forename: Hong-Yun
authors.initials: HY
authors.lastname: Tong
authors.forename: Xue-Wen
authors.initials: XW
authors.lastname: Hou
authors.forename: Amy
authors.initials: A
authors.lastname: Shelton
authors.forename: Mike
authors.initials: M
authors.lastname: Minton
authors.forename: Raymond
authors.initials: R
authors.lastname: Esquerra
authors.forename: Sanja
authors.initials: S
authors.lastname: Roje
authors.forename: Hanjo
authors.initials: H
authors.lastname: Hellmann
authors.forename: Zheng-Hui
authors.initials: ZH
authors.lastname: He
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D001219:Aspartate Aminotransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D013489:Suppression, Genetic; D014466:Ultraviolet Rays; D025101:Vitamin B 6 / Q000378:metabolism*
chemical_list: D029681:Arabidopsis Proteins; C543709:RUS1 protein, Arabidopsis; C543710:RUS2 protein, Arabidopsis; D025101:Vitamin B 6; D001219:Aspartate Aminotransferases