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Abiotic stress response in yeast and metal-binding ability of a type 2 metallothionein-like protein (PutMT2) from Puccinellia tenuiflora

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Abstract

Metallothioneins are low-molecular weight and cysteine-rich metal-binding proteins that play predominant cellular roles in the scavenging of reactive oxygen species and in mediating metal metabolism. To evaluate the role of a type-2 metallothionein-like gene from Puccinellia tenuiflora (PutMT2), the gene was over-expressed in yeast, and growth was assessed under a variety of abiotic stress conditions including peroxide (H2O2), salinity (NaCl and NaHCO3), and metal stress. PutMT2 overexpression in yeast improved the tolerance of cells to H2O2, NaCl, NaHCO3, Zn2+, Fe2+, Fe3+, Cd2+, Cr6+, and Ag+, but increased the sensitivity of cells to Mn2+, Co2+, Cu2+, and Ni2+ compared with control cells. PutMT2 was then expressed in Escherichia coli BL21as a glutathione S-transferase (GST) fusion protein (GST-PutMT2), and the metal-binding ability of GST-PutMT2 was analyzed and compared with GST alone using inductively coupled plasma atomic emission spectroscopy. Results showed that PutMT2 could bind to Cr, Cd, Co, Ag, Ba, Pb, Mn, Zn, Fe, Cu, P, Al, and Mg, but not Ni and As. There was no evidence to suggest that PutMT2 exhibited a specific or selective binding tendency to any individual metal ion. PutMT2 protein bound to Zn, Na, and Cu in vivo, perhaps with the highest affinity for Cu. Taken together, our results suggest that PutMT2 protein could play an important role in improving metal tolerance by metal binding in yeast. However, additional studies are required to confirm these results and to clarify the function of PutMT2 in transgenic plants.

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Acknowledgments

This work was supported by two Grants from Program for Changjiang Scholars and Innovative Research Team in University (PCSIRT) to Shenkui Liu and Program for New Century Excellent Talents in University (NECT-10-0314) to Xinxin Zhang.

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Authors and Affiliations

  1. Key Laboratory of Saline-alkali Vegetation Ecology Restoration in Oil Field (SAVER), Ministry of Education, Alkali Soil Natural Environmental Science Centre (ASNESC), Northeast Forestry University, Harbin, 150040, China

    Min Zhang, Shenkui Liu & Xinxin Zhang

  2. Asian Natural Environment Science Centre (ANESC), The University of Tokyo, Midori Cho 1-1-1, Nishitokyo City, Tokyo, 188-0002, Japan

    Tetsuo Takano

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  1. Min Zhang
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Correspondence to Xinxin Zhang.

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Zhang, M., Takano, T., Liu, S. et al. Abiotic stress response in yeast and metal-binding ability of a type 2 metallothionein-like protein (PutMT2) from Puccinellia tenuiflora . Mol Biol Rep 41, 5839–5849 (2014). https://doi.org/10.1007/s11033-014-3458-1

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