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Anchoring plant metallothioneins to the inner face of the plasma membrane of Saccharomyces cerevisiae cells leads to heavy metal accumulation
- Source :
- PLoS ONE, PLoS ONE, Vol 12, Iss 5, p e0178393 (2017)
- Publication Year :
- 2017
- Publisher :
- Public Library of Science (PLoS), 2017.
-
Abstract
- 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. © 2017 Ruta et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
- Subjects :
- 0106 biological sciences
0301 basic medicine
Cell Membranes
Arabidopsis
lcsh:Medicine
Yeast and Fungal Models
Heavy Metals
Toxicology
Pathology and Laboratory Medicine
Biochemistry
01 natural sciences
Green fluorescent protein
Medicine and Health Sciences
Fluorescence microscope
Toxins
Arabidopsis thaliana
Cloning, Molecular
lcsh:Science
Multidisciplinary
biology
Reverse Transcriptase Polymerase Chain Reaction
Plants
Complementary DNA
Nucleic acids
Chemistry
Experimental Organism Systems
Physical Sciences
Cellular Structures and Organelles
Research Article
Chemical Elements
DNA, Complementary
Forms of DNA
Arabidopsis Thaliana
Green Fluorescent Proteins
Toxic Agents
Saccharomyces cerevisiae
Brassica
Research and Analysis Methods
Saccharomyces
03 medical and health sciences
Model Organisms
Plant and Algal Models
Metals, Heavy
010608 biotechnology
Genetics
Molecular Biology Techniques
Molecular Biology
Myristoylation
Arabidopsis Proteins
lcsh:R
Cell Membrane
Organisms
Fungi
Biology and Life Sciences
Membrane Proteins
Cell Biology
DNA
biology.organism_classification
Yeast
Cytosol
030104 developmental biology
Membrane protein
lcsh:Q
Metallothionein
Cloning
Subjects
Details
- ISSN :
- 19326203
- Volume :
- 12
- Database :
- OpenAIRE
- Journal :
- PLOS ONE
- Accession number :
- edsair.doi.dedup.....2b990e1d47fe356c78f910490dfce419