NOTICE: The Editor-in-Chief and Publisher of Journal of Experimental Botany have taken the decision to retract the article related to this dataset due to concern over image manipulation and duplication. Please note, Dryad cannot verify whether the underlying data published is reliable for reuse. Visit this link to view the retraction notice and concerns raised: https://doi.org/10.1093/jxb/ery342
Members of the cation diffusion facilitator family have been identified in all kingdoms of life. They have been divided into three subgroups – Zn-CDF, Fe/Zn-CDF and Mn-CDF – based on their putative specificity to transported metal ions. The plant metal tolerance 6 (MTP6) proteins fall into the Fe/Zn-CDF subgroup, however their function in iron/zinc transport has not been confirmed yet. Here we characterize the MTP6 protein from cucumber. When expressed in yeast and protoplasts isolated from A. thaliana cells, CsMTP6 localized in mitochondria and contributed to the efflux of iron and manganese from the mitochondria. The immunolocalization of CsMTP6 in cucumber membranes confirmed that CsMTP6 is associated with mitochondria. The root expression and protein level of CsMTP6 were significantly up-regulated in conditions of iron deficiency and iron excess but were not affected by Mn availability. These results indicate that plant MTP6 proteins contribute to the distribution of iron and manganese between the cytosol and mitochondria of plant cells and are regulated by Fe to maintain the mitochondrial and cytosolic iron homeostasis under various Fe availability.
CsMTP6 expression data
Organ expression pattern of CsMTP6 in cucumber. Real-time expression analyses of the level of CsMTP6 transcript in the roots, hypocotyls, cotyledons, leaf petioles and leaves of 2-week-old cucumber seedlings growing in standard nutrition media. The CsMTP6 transcript levels relative to the constitutively expressed reference gene CACS were calculated (Lightcycler 480, Roche software) from the arithmetic means of ΔCp values obtained in three independent experiments. each experiment was performed in triplicate.
GDO activity in yeast data
Cytosolic gentisate dioxigenase (c-GDO) activity in the wild type (WT) or yeast mutant cells (mmt1/2) expressing c-GDO or c-GDO and cucumber CsMTP6 protein. c-GDO activity is expressed as nanomoles of substrate converted per minute per mg of protein. The data obtained come from the analysis of four individual yeast transformants.
Metal content in yeast data
Mitochondrial metals (Fe, Cu, Co, Zn, Mn, Cd) content in yeast transformed with empty vector or expressing CsMTP6. The yeast were grown in the presence of 100 microM concentration of each metal for 12 hours prior the preparation of mitochondria. metals content was determined by Atomic Absorption Spectrophotometer (AAS 3300, Perkin Elmer). The data are expressed in nmol x mg-1 protein and come from three separate experiments.
Metal content in mitochondria from yeast data.xls
Metal content in protoplasts data
Mitochondrial metals (Fe, Mn) content in protoplasts prepared from Arabidopsis thaliana suspension cells and transformed with empty vector pA7 or with the vector carrying cucumber CsMTP6 gene. Following transformation, protoplasts were incubated with 100 mcroM FeSO4-EDTA or 100 microM MnSO4 with slow agitation. Metal content was determined by AAS.The data are expressed in nmol of metal x mg-1 protein and were obtained in three separate experiments
Metal content in mitochondria of protoplasts data.xls
Supplementary Figures and Tables
PDF document with supplementary tables and figures:
Supplementary Table S1. Yeast strains used in this work;
Supplementary Fig. S1. Organ expression pattern of CsMTP6 in cucumber; Supplementary Fig. S2. ClustalW alignment of the CsMTP6 nucleic acid and CsMTP6 amino acid sequences;
Supplementary Fig. S3. Western blot analysis of mitochondria isolated from yeast expressing CsMTP6;
Supplementary Fig. S4. Localization of CsMTP6 in Δzrc1, K667 and Δsmf1 cells;
Supplementary Fig. S5. Effect of CsMTP6 expression on yeast sensitivity to Zn, Ni and Cd.
Supplementary data.pdf
