title: Acireductone dioxygenase 1 (ARD1) is an effector of the heterotrimeric G protein beta subunit in Arabidopsis.
abstract: Heterotrimeric G protein complexes are conserved from plants to mammals, but the complexity of each system varies. Arabidopsis thaliana contains one Gα, one Gβ (AGB1), and at least three Gγ subunits, allowing it to form three versions of the heterotrimer. This plant model is ideal for genetic studies because mammalian systems contain hundreds of unique heterotrimers. The activation of these complexes promotes interactions between both the Gα subunit and the Gβγ dimer with enzymes and scaffolds to propagate signaling to the cytoplasm. However, although effectors of Gα and Gβ are known in mammals, no Gβ effectors were previously known in plants. Toward identifying AGB1 effectors, we genetically screened for dominant mutations that suppress Gβ-null mutant (agb1-2) phenotypes. We found that overexpression of acireductone dioxygenase 1 (ARD1) suppresses the 2-day-old etiolated phenotype of agb1-2. ARD1 is homologous to prokaryotic and eukaryotic ARD proteins; one function of ARDs is to operate in the methionine salvage pathway. We show here that ARD1 is an active metalloenzyme, and AGB1 and ARD1 both control embryonic hypocotyl length by modulating cell division; they also may contribute to the production of ethylene, a product of the methionine salvage pathway. ARD1 physically interacts with AGB1, and ARD enzymatic activity is stimulated by AGB1 in vitro. The binding interface on AGB1 was deduced using a comparative evolutionary approach and tested using recombinant AGB1 mutants. A possible mechanism for AGB1 activation of ARD1 activity was tested using directed mutations in a loop near the substrate-binding site.
authors.affiliation: Department of Biology, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
authors.forename: Erin J
authors.initials: EJ
authors.lastname: Friedman
authors.forename: Helen X
authors.initials: HX
authors.lastname: Wang
authors.forename: Kun
authors.initials: K
authors.lastname: Jiang
authors.forename: Iva
authors.initials: I
authors.lastname: Perovic
authors.forename: Aditi
authors.initials: A
authors.lastname: Deshpande
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Pochapsky
authors.forename: Brenda R S
authors.initials: BR
authors.lastname: Temple
authors.forename: Stephanie N
authors.initials: SN
authors.lastname: Hicks
authors.forename: T Kendall
authors.initials: TK
authors.lastname: Harden
authors.forename: Alan M
authors.initials: AM
authors.lastname: Jones
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D002455:Cell Division / Q000502:physiology*; D049308:Dioxygenases / Q000235:genetics / Q000378:metabolism*; D005030:Ethylenes / Q000096:biosynthesis; D044387:GTP-Binding Protein beta Subunits / Q000235:genetics / Q000378:metabolism*; D005799:Genes, Dominant; D018546:Hypocotyl / Q000235:genetics / Q000378:metabolism*; D008715:Methionine / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D011485:Protein Binding; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins
chemical_list: C089590:AGB1 protein, Arabidopsis; D029681:Arabidopsis Proteins; D005030:Ethylenes; D044387:GTP-Binding Protein beta Subunits; D011994:Recombinant Proteins; C036216:ethylene; D008715:Methionine; D049308:Dioxygenases; C083734:aci-reductone oxidase (CO-forming)