Cloning of nicotianamine synthase genes from<i>Arabidopsis thaliana</i>

Suzuki, Kazuya; Higuchi, Kyoko; Nakanishi, Hiromi; Nishizawa, Naoko K.; Mori, Satoshi

doi:10.1080/00380768.1999.10414350

Cited by 58 publications

(47 citation statements)

References 36 publications

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“…6). Three MBP fusions of Arabidopsis NAS proteins (AtNAS1, AtNAS2, and AtNAS3) were also active in vitro (Suzuki et al, 1999). These results suggest that other MBP-NAS fusions should also exhibit NAS activity when produced in E. coli, supported by the finding that MBPZmNAS1 and MBP-ZmNAS3 were as active in vitro as MBP-HvNAS1.…”

Section: Inactive Zmnas2 In Spite Of the Duplicated Structuresupporting

confidence: 74%

“…NAS enzyme activity was assayed using the cell-free system modified by Higuchi et al (1994) and Suzuki et al (1999). One microgram of each fusion protein was added to the reaction buffer [50 mm Tris-HCl, 1 mm EDTA, 3 mm dithiothreitol, 10 m (p-amidinophenyl) methanesulfonyl fluoride (p-APSMF), and 10 m trans-epoxysuccinyl-l-leucylamido(4-guanidino) butane (E-64), pH 8.7] and was concentrated by ultrafiltration using a filter (Ultrafree C3LGC NMWL10000; Millipore, Bedford, MA).…”

Section: Assay Of Nas Activitymentioning

confidence: 99%

“…In these plants, NA has been implicated in the internal transport of metal ions (Scholz et al, 1992;Stephan and Scholz, 1993;Pich et al, 1994;Stephan et al, 1994). Since Higuchi et al (1999b) isolated the first NAS gene from barley, NAS genes have been isolated from barley again (Herbik et al, 1999), and from tomato (Lycopersicon esculentum; Ling et al, 1999), Arabidopsis (Suzuki et al, 1999), and rice (Oryza sativa; Higuchi et al, 2001). In this study, we focus on maize, an economically important cereal that is susceptible to Fe deficiency.…”

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Three Nicotianamine Synthase Genes Isolated from Maize Are Differentially Regulated by Iron Nutritional Status

et al. 2003

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Nicotianamine synthase (NAS) is an enzyme that is critical for the biosynthesis of the mugineic acid family of phytosiderophores in graminaceous plants, and for the homeostasis of metal ions in nongraminaceous plants. We isolated one genomic NAS clone, ZmNAS3, and two cDNA NAS clones, ZmNAS1 and ZmNAS2, from maize (Zea mays cv Alice). In agreement with the increased secretion of phytosiderophores with Fe deficiency, ZmNAS1 and ZmNAS2 were positively expressed only in Fe-deficient roots. In contrast, ZmNAS3 was expressed under Fe-sufficient conditions, and was negatively regulated by Fe deficiency. This is the first report describing down-regulation of NAS gene expression in response to Fe deficiency in plants, shedding light on the role of nicotianamine in graminaceous plants, other than as a precursor in phytosiderophore production. ZmNAS1-green fluorescent protein (sGFP) and ZmNAS2-sGFP were localized at spots in the cytoplasm of onion (Allium cepa) epidermal cells, whereas ZmNAS3-sGFP was distributed throughout the cytoplasm of these cells. ZmNAS1 and ZmNAS3 showed NAS activity in vitro, whereas ZmNAS2 showed none. Due to its duplicated structure, ZmNAS2 was much larger (65.8 kD) than ZmNAS1, ZmNAS3, and previously characterized NAS proteins (30-38 kD) from other plant species. We reveal that maize has two types of NAS proteins based on their expression pattern and subcellular localization.To acquire Fe, graminaceous plants secrete Fe chelators, known as mugineic-acid family phytosiderophores (MAs). MAs dissolve Fe in the rhizosphere, followed by reabsorption of the Fe(III)-MA complexes through YS1 transporters in the plasma membrane (Takagi, 1976; Curie et al., 2001). Only graminaceous plants use the MA mechanism of acquiring Fe(III), classified as the Strategy II mechanism (Marschner et al., 1986). Fe deficiency is a problem in crop production worldwide, especially in calcareous soils, where Fe is sparingly soluble due to the high soil pH. The ability of graminaceous plants to tolerate Fe deficiency is thought to depend on the quantity of MAs secreted during Fe deficiency (Takagi, 1976;Mori et al., , 1988Rö mheld, 1987;Kawai et al., 1988;Mihashi and Mori, 1989;Singh et al., 1993). The biosynthetic pathways of MAs (Fig. 1A) have been determined Kawai et al., 1988;Shojima et al., 1990;Ma et al., 1999), and almost all the genes involved have been isolated in our laboratory (Higuchi et al., 1999b;Takahashi et al., 1999;Nakanishi et al., 2000;Kobayashi et al., 2001). NAS is a key enzyme in MA biosynthesis, catalyzing the trimerization of SAM into one molecule of NA (Higuchi et al., 1999b). NAS activity in graminaceous plants is well correlated with tolerance to Fe deficiency. In maize (Zea mays), a plant susceptible to Fe deficiency, NAS activity is very low (Higuchi et al., 1996a), and maize secretes lower amounts of MAs than barley or oat (Avena sativa), cereals that are tolerant of low Fe supply (Rö mheld, 1987;Lytle and Jolley, 1991). NAS is also important for growth in nongraminaceous plants, which do not ...

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Section: Inactive Zmnas2 In Spite Of the Duplicated Structuresupporting

confidence: 74%

Section: Assay Of Nas Activitymentioning

confidence: 99%

mentioning

confidence: 99%

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Three Nicotianamine Synthase Genes Isolated from Maize Are Differentially Regulated by Iron Nutritional Status

et al. 2003

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“…These chelators are either enzymatically synthesized small molecular weight compounds, such as nicotianamine (NA) 1 and phytochelatin, or proteins, such as metallothionein (MT) and metal chaperones. Four NA synthase genes, NAS1-3 (14), two phytochelatin synthase (PCS) genes (15), and seven MT genes (16 -18) have been identified. AtCCH, a homologue to the copper chaperone Atx1p of Saccharomyces cerevisiae, plays a role in intracellular copper transport (19,20), while the function of 29 additional proteins containing an Atx1p-like heavy metal binding site (HMA) is unknown (21,22).…”

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confidence: 99%

Expression Profiles of Arabidopsis thaliana in Mineral Deficiencies Reveal Novel Transporters Involved in Metal Homeostasis

Wintz

Fox

et al. 2003

Journal of Biological Chemistry

379

330

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Plants directly assimilate minerals from the environment and thus are key for acquisition of metals by all subsequent consumers. Limited bio-availability of copper, zinc and iron in soil decreases both the agronomic productivity and the nutrient quality of crops. Understanding the molecular mechanisms underlying metal homeostasis in plants is a prerequisite to optimizing plant yield and metal nutrient content. To absorb and maintain a balance of potentially toxic metal ions, plants utilize poorly understood mechanisms involving a large number of membrane transporters and metalbinding proteins with overlapping substrate specificities and complex regulation. To better understand the function and the integrated regulation, we analyzed in Arabidopsis the expression patterns in roots and in leaves of 53 genes coding for known or potential metal transporters, in response to copper, zinc, and iron deficiencies in Arabidopsis. Comparative analysis of gene expression profiles revealed specific transcriptional regulation by metals of the genes contrasting with the known wide substrate specificities of the encoded transporters. Our analysis suggested novel transport roles for several gene products and we used functional complementation of yeast mutants to correlate specific regulation by metals with transport activity. We demonstrate that two ZIP genes, ZIP2 and ZIP4, are involved in copper transport. We also present evidence that AtOPT3, a member of the oligopeptide transporter gene family with significant similarities to the maize iron-phytosiderophore transporter YS1, is regulated by metals and heterologous expression AtOPT3 can rescue yeast mutants deficient in metal transport.All organisms require metal prosthetic groups for their unique catalytic and structural properties. In proteins, copper and iron catalyze reduction-oxidation reactions, while zinc plays an essential structural or enzymatic role. Yet most metal ions are very reactive and can be toxic to cells when present in excess. Thus, it is important for organisms to maintain adequate levels of metals in tight homeostasis using complex and often evolutionarily conserved mechanisms for the uptake and transport of low solubility metals and storage of metal ions in a non-toxic form. Despite rapid progress in recent years of our understanding of metal homeostasis in yeast, (1) our knowledge of metal metabolism in plants is still rudimentary (2).A large number of cation transporters potentially involved in metal ion homeostasis have been identified on the genome of the model plant Arabidopsis thaliana (3). Several members of the 15 ZIP gene family (4) and of the 6 NRAMP family of transporters (3) have been characterized and shown to be involved in metal uptake and transport in plants (5-10). ABC transporters (11) and P-type ATPase pumps (12, 13) are also known to be involved in metal ions trafficking metals into organelles. Since metals are cytotoxic as free ions, they are chelated for both intracellular storage and long-distance transport. These chelators are eith...

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“…Previously, we confirmed the enzyme activities of AtNAS1-3 in vitro. 40 To gain further insight into physiological functions of NASs, we analyzed subcellular localization of AtNAS1-4 by transient expression of sGFP fusions in tobacco BY-2 cells (Fig. 2).…”

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Rice nicotianamine synthase localizes to particular vesicles for proper function

Nozoye

Tsunoda

Nagasaka

et al. 2014

Plant Signaling & Behavior

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Cloning of nicotianamine synthase genes fromArabidopsis thaliana

Cited by 58 publications

References 36 publications

Three Nicotianamine Synthase Genes Isolated from Maize Are Differentially Regulated by Iron Nutritional Status

Three Nicotianamine Synthase Genes Isolated from Maize Are Differentially Regulated by Iron Nutritional Status

Expression Profiles of Arabidopsis thaliana in Mineral Deficiencies Reveal Novel Transporters Involved in Metal Homeostasis

Rice nicotianamine synthase localizes to particular vesicles for proper function

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