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Characterization of an Arabidopsis enzyme family that conjugates amino acids to indole-3-acetic acid.
- Source :
-
The Plant cell [Plant Cell] 2005 Feb; Vol. 17 (2), pp. 616-27. Date of Electronic Publication: 2005 Jan 19. - Publication Year :
- 2005
-
Abstract
- Substantial evidence indicates that amino acid conjugates of indole-3-acetic acid (IAA) function in auxin homeostasis, yet the plant enzymes involved in their biosynthesis have not been identified. We tested whether several Arabidopsis thaliana enzymes that are related to the auxin-induced soybean (Glycine max) GH3 gene product synthesize IAA-amino acid conjugates. In vitro reactions with six recombinant GH3 enzymes produced IAA conjugates with several amino acids, based on thin layer chromatography. The identity of the Ala, Asp, Phe, and Trp conjugates was verified by gas chromatography-mass spectrometry. Insertional mutations in GH3.1, GH3.2, GH3.5, and GH3.17 resulted in modestly increased sensitivity to IAA in seedling root. Overexpression of GH3.6 in the activation-tagged mutant dfl1-D did not significantly alter IAA level but resulted in 3.2- and 4.5-fold more IAA-Asp than in wild-type seedlings and mature leaves, respectively. In addition to IAA, dfl1-D was less sensitive to indole-3-butyric acid and naphthaleneacetic acid, consistent with the fact that GH3.6 was active on each of these auxins. By contrast, GH3.6 and the other five enzymes tested were inactive on halogenated auxins, and dfl1-D was not resistant to these. This evidence establishes that several GH3 genes encode IAA-amido synthetases, which help to maintain auxin homeostasis by conjugating excess IAA to amino acids.
- Subjects :
- Arabidopsis enzymology
Arabidopsis Proteins metabolism
Gene Expression Regulation, Plant
Homeostasis
Multienzyme Complexes genetics
Multienzyme Complexes metabolism
Mutagenesis, Insertional
Phylogeny
Plant Roots growth & development
Plant Roots metabolism
Recombinant Proteins genetics
Recombinant Proteins metabolism
Substrate Specificity
Amino Acids metabolism
Arabidopsis genetics
Arabidopsis Proteins genetics
Indoleacetic Acids metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1040-4651
- Volume :
- 17
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- The Plant cell
- Publication Type :
- Academic Journal
- Accession number :
- 15659623
- Full Text :
- https://doi.org/10.1105/tpc.104.026690