title: AtNOS/AtNOA1 is a functional Arabidopsis thaliana cGTPase and not a nitric-oxide synthase.
abstract: AtNOS1 was previously identified as a potential nitric-oxide synthase (NOS) in Arabidopsis thaliana, despite lack of sequence similarity to animal NOSs. Although the dwarf and yellowish leaf phenotype of Atnos1 knock-out mutant plants can be rescued by treatment with exogenous NO, doubts have recently been raised as to whether AtNOS1 is a true NOS. Moreover, depending on the type of physiological responses studied, Atnos1 is not always deficient in NO induction and/or detection, as previously reported. Here, we present experimental evidence showing that AtNOS1 is unable to bind and oxidize arginine to NO. These results support the argument that AtNOS1 is not a NOS. We also show that the renamed NO-associated protein 1 (AtNOA1) is a member of the circularly permuted GTPase family (cGTPase). AtNOA1 specifically binds GTP and hydrolyzes it. Complementation experiments of Atnoa1 mutant plants with different constructs of AtNOA1 show that GTP hydrolysis is necessary but not sufficient for the physiological function of AtNOA1. Mutant AtNOA1 lacking the C-terminal domain, although retaining GTPase activity, failed to complement Atnoa1, suggesting that this domain plays a crucial role in planta. cGTPases appear to be RNA-binding proteins, and the closest homolog of AtNOA1, the Bacillus subtilis YqeH, has been shown to participate in ribosome assembly and stability. We propose a similar function for AtNOA1 and discuss it in the light of its potential role in NO accumulation and plant development.
authors.affiliation: Boyce Thompson Institute for Plant Research, Ithaca, New York 14853, USA.
authors.forename: Magali
authors.initials: M
authors.lastname: Moreau
authors.forename: Gyu In
authors.initials: GI
authors.lastname: Lee
authors.forename: Yongzeng
authors.initials: Y
authors.lastname: Wang
authors.forename: Brian R
authors.initials: BR
authors.lastname: Crane
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Klessig
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology*; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000502:physiology*; D001412:Bacillus subtilis / Q000378:metabolism; D020558:GTP Phosphohydrolases / Q000737:chemistry*; D006868:Hydrolysis; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase / Q000737:chemistry / Q000378:metabolism* / Q000502:physiology; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012270:Ribosomes / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D019076:Transgenes
chemical_list: D029681:Arabidopsis Proteins; D009569:Nitric Oxide; C521947:NOA1 protein, Arabidopsis; D019001:Nitric Oxide Synthase; D020558:GTP Phosphohydrolases
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