Expression of the rgMT gene, encoding for a rice metallothionein-like protein in Saccharomyces cerevisiae and Arabidopsis thaliana

Jin, Shumei; Sun, Dan; Wang, Ji; Li, Ying; Wang, Xinwang; Liu, Shenkui

doi:10.1007/s12041-014-0430-8

Cited by 16 publications

(11 citation statements)

References 49 publications

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“…Plant MTs are also heavily involved in both metal metabolism and detoxification . Due to the difficulty in isolating native MT proteins from plants, their metal‐binding properties are generally characterized by heterologous expression in bacteria . In this study, heterologous expression of CsMTL3 led to increased metal tolerance and metal ion accumulation, especially for Cu and Cd (Figs and ).…”

Section: Discussionmentioning

confidence: 84%

“…MTs are highly enriched in cysteine (Cys) residues [6,7] with metal-binding motifs (Cys-Cys, Cys-X-Cys, or Cys-X-X-Cys) that provide sulfhydryl ligands for coordinating bivalent metal ions [8,9]. They have been grouped into different classes according to the arrangement of their Cys residues [6,[10][11][12][13][14][15][16]. MTs that contain 20 highly conserved Cys residues were classified as class I MTs and are generally found in vertebrates [17,18].…”

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confidence: 99%

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Characterization analysis and heavy metal‐binding properties of CsMTL3 in Escherichia coli

Liu

et al. 2018

FEBS Open Bio

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Members of the metallothionein ( MT ) superfamily are involved in coordinating transition metal ions. In plants, MT family members are characterized by their arrangement of Cys residues. In this study, one member of the MT superfamily, Cs MTL 3 , was characterized from a complementary DNA ( cDNA ) library from young cucumber fruit; Cs MTL 3 is predicted to encode a 64 amino acid protein with a predicted molecular mass of 6.751 kDa. Phylogenetic analysis identified it as a type 3 family member as the arrangement of N‐terminal Cys residues was different from that of MT‐like 2. Heterologous expression of Cs MTL 3 in Escherichia coli improved their heavy metal tolerance, particularly to Cd 2+ and Cu 2+ , and led to increased uptake of Cd 2+ and Cu 2+ ; increased uptake was also observed for cells expressing Arabidopsis thaliana metallothionein 3 (At MT 3) and phytochelatin‐like ( PCL ), with greatest uptake in PCL ‐expressing cells. These findings demonstrate that Cs MTL 3 can improve metal tolerance, especially for Cd 2+ ions, when heterologously expressed in E. coli , and suggest that the composition and arrangement of N‐terminal Cys residues are associated with binding capacity and preference for different metal ions.

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Section: Discussionmentioning

confidence: 84%

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confidence: 99%

Characterization analysis and heavy metal‐binding properties of CsMTL3 in Escherichia coli

Liu

et al. 2018

FEBS Open Bio

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“…Based on their arrangement of Cys residues, MTs can be divided into three classes (I, II, and III) [1,7,10,11,12,13,14,15]. Class I MTs, which contain 20 highly conserved Cys residues, are widely distributed in vertebrates [16,17]; Class II MTs, which have a flexible arrangement of Cys residues, are encoded by a family of genes that are ubiquitous in plants, fungi, animals, and cyanobacteria [1,12,15,18].…”

Section: Introductionmentioning

confidence: 99%

“…For example, Arabidopsis MT1 knock-down lines exhibit Cd sensitivity, and transgenic tobacco plants heterologously expressing Arabidopsis MT genes are resistant to Cd toxicity [12]. In addition, some plant MT2 genes have also been heterologously expressed in various organisms, and were found to impart increased tolerance to metals in E. coli [4,13,26,31]. …”

Section: Introductionmentioning

confidence: 99%

Cucumber Metallothionein-Like 2 (CsMTL2) Exhibits Metal-Binding Properties

Pan

Zhai

et al. 2016

Genes

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We identified a novel member of the metallothionein (MT) family, Cucumis sativus metallothionein-like 2 (CsMTL2), by screening a young cucumber fruit complementary DNA (cDNA) library. The CsMTL2 encodes a putative 77-amino acid Class II MT protein that contains two cysteine (Cys)-rich domains separated by a Cys-free spacer region. We found that CsMTL2 expression was regulated by metal stress and was specifically induced by Cd2+ treatment. We investigated the metal-binding characteristics of CsMTL2 and its possible role in the homeostasis and/or detoxification of metals by heterologous overexpression in Escherichia coli cells. Furthermore, we produced a deletion mutant form of the protein, CsMTL2m, that contained the two Cys-rich clusters but lacked the spacer region, in E. coli. We compared the metal-binding properties of CsMTL2 with those of CsMTL2m, the β domain of human metallothionein-like protein 1 (HsMTXb), and phytochelatin-like (PCL) heterologously expressed in E. coli using metal-binding assays. We found that E. coli cells expressing CsMTL2 accumulated the highest levels of Zn2+ and Cd2+ of the four transformed cell types, with levels being significantly higher than those of control cells containing empty vector. E. coli cells expressing CsMTL2 had a higher tolerance for cadmium than for zinc ions. These findings show that CsMTL2 improves metal tolerance when heterologously expressed in E. coli. Future studies should examine whether CsMTL2 improves metal tolerance in planta.

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“…Saccharomyces cerevisiae has often been used to express plant methallothioneins (MTs) either for functional elucidation through complementation studies [24, 28–31] or for their metal-binding abilities [32, 42–45]. While most of the studies concerning heterologous expression of MTs in yeast focus on canonical MT substrates, our work encompassed a greater variety of both natural and non-canonical MT substrates, all in parallel settings.…”

Section: Discussionmentioning

confidence: 99%

Anchoring plant metallothioneins to the inner face of the plasma membrane of Saccharomyces cerevisiae cells leads to heavy metal accumulation

et al. 2017

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In this study we engineered yeast cells armed for heavy metal accumulation by targeting plant metallothioneins to the inner face of the yeast plasma membrane. Metallothioneins (MTs) are cysteine-rich proteins involved in the buffering of excess metal ions, especially Cu(I), Zn(II) or Cd(II). The cDNAs of seven Arabidopsis thaliana MTs (AtMT1a, AtMT1c, AtMT2a, AtMT2b, AtMT3, AtMT4a and AtMT4b) and four Noccaea caerulescens MTs (NcMT1, NcMT2a, NcMT2b and NcMT3) were each translationally fused to the C-terminus of a myristoylation green fluorescent protein variant (myrGFP) and expressed in Saccharomyces cerevisiae cells. The myrGFP cassette introduced a yeast myristoylation sequence which allowed directional targeting to the cytosolic face of the plasma membrane along with direct monitoring of the intracellular localization of the recombinant protein by fluorescence microscopy. The yeast strains expressing plant MTs were investigated against an array of heavy metals in order to identify strains which exhibit the (hyper)accumulation phenotype without developing toxicity symptoms. Among the transgenic strains which could accumulate Cu(II), Zn(II) or Cd(II), but also non-canonical metal ions, such as Co(II), Mn(II) or Ni(II), myrGFP-NcMT3 qualified as the best candidate for bioremediation applications, thanks to the robust growth accompanied by significant accumulative capacity.

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Expression of the rgMT gene, encoding for a rice metallothionein-like protein in Saccharomyces cerevisiae and Arabidopsis thaliana

Cited by 16 publications

References 49 publications

Characterization analysis and heavy metal‐binding properties of CsMTL3 in Escherichia coli

Characterization analysis and heavy metal‐binding properties of CsMTL3 in Escherichia coli

Cucumber Metallothionein-Like 2 (CsMTL2) Exhibits Metal-Binding Properties

Anchoring plant metallothioneins to the inner face of the plasma membrane of Saccharomyces cerevisiae cells leads to heavy metal accumulation

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