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Originally published In Press as doi:10.1074/jbc.M111955200 on March 28, 2002

J. Biol. Chem., Vol. 277, Issue 23, 20446-20452, June 7, 2002
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Characterization of a Family of IAA-Amino Acid Conjugate Hydrolases from Arabidopsis*

Sherry LeClereDagger §, Rosie TellezDagger , Rebekah A. RampeyDagger , Seiichi P. T. MatsudaDagger ||, and Bonnie BartelDagger **

From the Departments of Dagger  Biochemistry and Cell Biology and || Chemistry, Rice University, Houston, Texas 77005

The mechanisms by which plants regulate levels of the phytohormone indole-3-acetic acid (IAA) are complex and not fully understood. One level of regulation appears to be the synthesis and hydrolysis of IAA conjugates, which function in both the permanent inactivation and temporary storage of auxin. Similar to free IAA, certain IAA-amino acid conjugates inhibit root elongation. We have tested the ability of 19 IAA-L-amino acid conjugates to inhibit Arabidopsis seedling root growth. We have also determined the ability of purified glutathione S-transferase (GST) fusions of four Arabidopsis IAA-amino acid hydrolases (ILR1, IAR3, ILL1, and ILL2) to release free IAA by cleaving these conjugates. Each hydrolase cleaves a subset of IAA-amino acid conjugates in vitro, and GST-ILR1, GST-IAR3, and GST-ILL2 have Km values that suggest physiological relevance. In vivo inhibition of root elongation correlates with in vitro hydrolysis rates for each conjugate, suggesting that the identified hydrolases generate the bioactivity of the conjugates.

* This work was supported by the National Institutes of Health (R29 GM54749) and the Robert A. Welch Foundation (C-1309 and C-1323).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AY065996.

§ Supported in part by National Institutes of Health Biotechnology Training Grant T32 GM08362.

Supported in part by Houston Livestock Show and Rodeo scholarships.

** To whom correspondence should be addressed: Dept. of Biochemistry and Cell Biology, Rice University, 6100 S. Main St., Houston, TX 77005. Tel.: 713-348-5602; Fax: 713-348-5154; E-mail: bartel@rice.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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