The gene for indole-3-acetyl-L-aspartic acid hydrolase from Enterobacter agglomerans : molecular cloning, nucleotide sequence, and expression in Escherichia coli

Chou, Jyh Ching; Mulbry, Walter; Cohen, Jerry D.

doi:10.1007/s004380050802

Cited by 35 publications

(15 citation statements)

References 19 publications

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“…6). In agreement with this lack oí activity is the fact that the amino acid sequence of ^4/AMIl is unrelated to the reported sequences of auxin amide-conjugate hydrolases from the bacterium Enterobacter agglomerans (Chou et al, 1998) and Arabidopsis thaliana (Davies et al, 1999;LeClere et al, 2002).…”

Section: Discussionmentioning

confidence: 76%

Molecular cloning and characterization of an amidase from Arabidopsis thaliana capable of converting indole-3-acetamide into the plant growth hormone, indole-3-acetic acid

2003

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Acylamidohydrolases from higher plants have not been characterized or cloned so far. AtPsMll is the first member of this enzyme family from a higher plant and was identified in the genome of Arabidopsis thaliana based on sequence homology with the catalyticdomain sequence of bacterial acylamidohydrolases, particularly those that exhibit indole-3-acetamide amidohydrolase activity. AtAMll polypeptide and mRNA are present in leaf tissues, as shown by immunoblotting and RT-PCR, respectively. AtAMll was expressed from its cDNA in enzymatically active form and exhibits substrate specificity for indole-3-acetamide, but also some activity against L-asparagine. The recombinant enzyme was characterized further. The results show that higher plants have acylamidohydrolases with properties similar to the enzymes of certain plant-associated bacteria such as Agrobacterium-, Pseudomonasand Rhodococcus-spQCÍQS, in which these enzymes serve to synthesize the plant growth hormone, indole-3-acetic acid, utilized by the bacteria to colonize their host plants. As Índole-3-acetamide is a native metabolite in Arabidopsis thaliana, it can no longer be ruled out that one pathway for the biosynthesis of indole-3-acetic acid involves indole-3-acetamide-hydrolysis by AtPsMll.

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Section: Discussionmentioning

confidence: 76%

Molecular cloning and characterization of an amidase from Arabidopsis thaliana capable of converting indole-3-acetamide into the plant growth hormone, indole-3-acetic acid

2003

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“…Two of these mutants, ilr1 and iar3, are defective in IAA-amino acid conjugate hydrolases (Bartel and Fink, 1995;Davies et al, 1999). The ILR1 and IAR3 enzymes hydrolyze IAA conjugates in vitro (Bartel and Fink, 1995;Davies et al, 1999;LeClere et al, 2002) and are similar to members of the M40 class of bacterial carboxypeptidases (Barrett et al, 2003) that cleave a variety of small molecules, including IAA-Asp (Chou et al, 1996(Chou et al, , 1998, benzoylglycine (Hani and Chan, 1995), acetylated amino acids (Sakanyan et al, 1993), and secreted human odorant precursors (Natsch et al, 2003). Degenerate PCR and sequence similarity searches revealed five additional ILR1-like genes (ILL1, ILL2, ILL3, ILL5, and ILL6) in the Arabidopsis amidohydrolase family (Davies et al, 1999;LeClere et al, 2002).…”

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confidence: 99%

A Family of Auxin-Conjugate Hydrolases That Contributes to Free Indole-3-Acetic Acid Levels during Arabidopsis Germination

et al. 2004

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Auxins are hormones important for numerous processes throughout plant growth and development. Plants use several mechanisms to regulate levels of the auxin indole-3-acetic acid (IAA), including the formation and hydrolysis of amide-linked conjugates that act as storage or inactivation forms of the hormone. Certain members of an Arabidopsis amidohydrolase family hydrolyze these conjugates to free IAA in vitro. We examined amidohydrolase gene expression using northern and promoterb-glucuronidase analyses and found overlapping but distinct patterns of expression. To examine the in vivo importance of auxin-conjugate hydrolysis, we generated a triple hydrolase mutant, ilr1 iar3 ill2, which is deficient in three of these hydrolases. We compared root and hypocotyl growth of the single, double, and triple hydrolase mutants on IAA-Ala, IAA-Leu, and IAAPhe. The hydrolase mutant phenotypic profiles on different conjugates reveal the in vivo activities and relative importance of ILR1, IAR3, and ILL2 in IAA-conjugate hydrolysis. In addition to defective responses to exogenous conjugates, ilr1 iar3 ill2 roots are slightly less responsive to exogenous IAA. The triple mutant also has a shorter hypocotyl and fewer lateral roots than wild type on unsupplemented medium. As suggested by the mutant phenotypes, ilr1 iar3 ill2 imbibed seeds and seedlings have lower IAA levels than wild type and accumulate IAA-Ala and IAA-Leu, conjugates that are substrates of the absent hydrolases. These results indicate that amidohydrolases contribute free IAA to the auxin pool during germination in Arabidopsis.Although auxins are essential regulators of many aspects of plant growth and development, our understanding of how levels of this hormone are controlled remains incomplete. One component of auxin homeostasis is conjugation of the auxin indole-3-acetic acid (IAA) to different moieties, including esterification to sugars and amide linkage to amino acids and peptides. IAA-Leu, IAA-Ala, IAA-Asp, IAA-Glu, and IAA-Glc have been identified in Arabidopsis seedlings (Tam et al., 2000;Kowalczyk and Sandberg, 2001). Several IAA-peptide conjugates have been identified in bean seeds (Bialek and Cohen, 1986;Walz et al., 2002) and Arabidopsis (Walz et al., 2002); in fact, IAApeptide conjugates are the major IAA conjugates in Arabidopsis seeds Park and Cohen, 2003). Different IAA conjugates apparently have specific functions in plants, such as storage, transport, or inactivation of IAA (Cohen and Bandurski, 1982;Bartel et al., 2001). In general, endogenous IAA conjugates that are biologically active and hydrolyzed in plants may function as auxin storage forms, whereas conjugates inactive in bioassays may have roles in IAA degradation Ljung et al., 2002).Several Arabidopsis screens have uncovered IAAamino acid conjugate-resistant mutants that help to delineate conjugate functions (Bartel and Fink, 1995;Campanella et al., 1996;Davies et al., 1999;Lasswell et al., 2000;Magidin et al., 2003;LeClere et al., 2004). Two of these mutants, ilr1 and iar3, are defective...

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“…New insights into the structure-function relationship of the putative PpIAR3 would add to our understanding of the evolution of all auxin amidohydrolases. So far, modeling and subsequent analysis of mutant enzymes has been carried out for a bacterial auxin conjugate hydrolase specific for IAA-Asp, [50] and now a few studies on plant enzyme counterparts have been added to the picture, [42,58] following the publication of the crystal structure of Arabidopsis ILL2 (an IAA specific auxin conjugate hydrolase, Fig. 1.…”

Section: Structure and Biochemistrymentioning

confidence: 99%

“…Interestingly, in this family of hydrolases, several ones were found which displayed activity towards IAA-Aspartate, [43] earlier described only as a substrate for bacterial IAA conjugate hydrolases. [50] Even though IAR3 enzyme was classified as an IAA amidohydrolase based on its initially examined in vitro activity against IAA conjugates, [51] more recently it was shown it has a dual role participating also in jasmonic acid (JA) homeostasis. [52,53] These results suggest that IAR3/JIH1 from Nicotiana attenuata [52] and Arabidopsis [53] is one of the major players of auxin-jasmonate crosstalk in plant stress responses.…”

Section: Auxin Amidohydrolases: Discoverymentioning

confidence: 99%

“…[61] Based on the sequence alignment of bacterial representatives of the M20 family [62] and the plant amidohydrolases, a few conserved motifs are present. [50,59] Promiscuity of the auxin amidohydrolases to accept structurally different substrates into the active site cavity [41−43] has probably arisen from the changes in the amino acid sequence of the active site cavity and/or in the conserved structural motifs present in all representatives of the M20 family, which lead to a fine tuning of the ability of an ancestor of auxin amidohydrolases to cleave various substrates across different families of land plants. [3] Quite possibly, the ancestor was a peptidase which was able to cleave a structurally related dipeptide, Trp-Asp as a substrate.…”

Section: Structure and Biochemistrymentioning

confidence: 99%

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Auxin Amidohydrolases – From Structure to Function: Revisited

Smolko¹,

Ludwig‐Müller²,

Salopek‐Sondi³

2018

Croat. Chem. Acta

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The control of plant growth and development is a well-coordinated process between exogenous and endogenous signals. Auxins are plant hormones belonging to the endogenous signals, which control a vast array of different processes. While auxins are growth promoting at low concentrations, higher levels are often inhibitory. Therefore, the tight control of auxin concentrations in a given plant tissue is essential. Among several processes that participate in auxin homeostasis, we focused herein on the process of reversible auxin conjugation that considers the synthesis of inactive auxin conjugates, which can be hydrolyzed back to the active form by so called auxin conjugate hydrolases. Although these proteins have been known for quite some time, their role in plants is still not clear, especially since novel hydrolases with different substrate specificities have been isolated. Thus, we have revisited the knowledge about auxin hydrolases, from their structure and biochemistry to the role in plant development and in dealing with unfavorable climate conditions.

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The gene for indole-3-acetyl-L-aspartic acid hydrolase from Enterobacter agglomerans : molecular cloning, nucleotide sequence, and expression in Escherichia coli

Cited by 35 publications

References 19 publications

Molecular cloning and characterization of an amidase from Arabidopsis thaliana capable of converting indole-3-acetamide into the plant growth hormone, indole-3-acetic acid

Molecular cloning and characterization of an amidase from Arabidopsis thaliana capable of converting indole-3-acetamide into the plant growth hormone, indole-3-acetic acid

A Family of Auxin-Conjugate Hydrolases That Contributes to Free Indole-3-Acetic Acid Levels during Arabidopsis Germination

Auxin Amidohydrolases – From Structure to Function: Revisited

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