DOI: 10.5061/dryad.306sf0b
title: Data from: Metal tolerance protein MTP6 affects mitochondrial iron and manganese homeostasis in cucumber

description: NOTICE: The Editor-in-Chief and Publisher of Journal of
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 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.
description: CsMTP6 expression
 dataOrgan 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
 dataCytosolic 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
 dataMitochondrial 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.xlsMetal content in
 protoplasts dataMitochondrial 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
 experimentsMetal content in mitochondria
 of protoplasts data.xlsSupplementary Figures
 and TablesPDF 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