Journal Article

Molecular Cloning and Characterization of Aldehyde Oxidases in <i>Arabidopsis thaliana</i>

Hiroyuki Sekimoto, Mitsunori Seo, Naoto Kawakami, Teruya Komano, Sophie Desloire, Sylviane Liotenberg, Annie Marion-Poll, Michel Caboche, Yuji Kamiya and Tomokazu Koshiba

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 39, issue 4, pages 433-442
Published in print April 1998 | ISSN: 0032-0781
Published online April 1998 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/oxfordjournals.pcp.a029387
Molecular Cloning and Characterization of Aldehyde Oxidases in Arabidopsis thaliana

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Using degenerate primers designed by deduced amino acid sequences of known aldehyde oxidases (AO) from maize and bovine, two independent cDNA fragments were amplified by reverse transcription-polymerase chain reaction (PCR). The two corresponding full-length cDNAs (atAO-1 and atAO-2; 4,484 and 4,228 bp long, respectively) were cloned by screening the Arabidopsis cDNA library followed by rapid amplification of cDNA end-PCR. These cDNAs are highly homologous at both the nucleotide and amino acid sequence levels, and the deduced amino acid sequences showed high similarity with those of maize and tomato AOs. They contain consensus sequences for two iron-sulfur centers and a molybdenum cofactor (MoCo)-binding domain. In addition, another cDNA having a sequence similar to that of the cDNAs was screened (atAO-3; 3,049 bp), and a putative AO gene (AC002376) was reported on chromosome 1, which (atAO-4) was distinct from, but very similar to, the above three AOs. atAO-1, 2, 3, and 4 were physically mapped on chromosomes 5, 3, 2 and 1, respectively. These data indicate that there is an AO multigene family in Arabidopsis. atAO-1 protein was shown to be highly similar to one of the maize AOs in respect to a region thought to be involved in determination of substrate specificity, suggesting that they might encode a similar type of AO, which could efficiently oxidize indole-3-acetaldehyde to indole-3-acetic acid (IAA). atAO-1 and atAO-2 genes were expressed at higher levels in lower hypo-cotyls and roots of the wild-type seedlings, while atAO-3 was slightly higher in cotyledons and upper hypocotyls. The expression of atAO-1 was more abundant in the seedlings of an IAA overproducing mutant (superrootl; surf) than in those of wild type. atAO-2 and atAO-3 transcripts were rather evenly distributed in these seedlings. A possible involvement of atAO genes in phytohormone biosynthesis in Arabidopsis is discussed.

Keywords: Abcisic acid; Aldehyde oxidase (EC 1.2.3.1); Arabidopsis thaliana; Indole-3-acetic acid (IAA); superrootl (surl) mutant

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Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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