The family of SMF metal ion transporters in yeast cells

A. Cohen, H. Nelson, N. Nelson

Research output: Contribution to journalArticlepeer-review

Abstract

Metal ions are vital for all organisms, and metal ion transporters play a crucial role in maintaining their homeostasis. The yeast (Saccharomyces cerevisiae) Smf transporters and their homologs in other organisms have a central role in the accumulation of metal ions and their distribution in different tissues and cellular organelles. In this work we generated null mutations in each individual SMF gene in yeast as well as in all combinations of the genes. Each null mutation exhibited sensitivity to metal ion chelators at different concentrations. The combination of null mutants ΔSMF1 + ΔSMF2 and the triple null mutant Δ3SMF failed to grow on medium buffered at pH 8 and 7.5, respectively. Addition of 5 μM copper or 25 μM manganese alleviated the growth arrest at the high pH or in the presence of the chelating agent. The transport of manganese was analyzed in the triple null mutant and in this mutant expressing each Smf protein. Although overexpression of Smf1p and Smf2p resulted in uptake that was higher than wild type cells, the expression of Smf3p gave no significant uptake above that of the triple mutant Δ3SMF. Western analysis with antibody against Smf3p indicated that this transporter does not reach the plasma membrane and may function at the Golgi or post-Golgi complexes. The iron uptake resulting from expression of Smf1p and Smf2p was analyzed in a mutant in which its iron transporters FET3 and FET4 were inactivated. Overexpression of Smflp gave rise to a significant iron uptake that was sensitive to the sodium concentrations in the medium. We conclude that the Smf proteins play a major role in copper and manganese homeostasis and, under certain circumstances, Smf1p may function in iron transport into the cells.

Original languageEnglish
Pages (from-to)33388-33394
Number of pages7
JournalJournal of Biological Chemistry
Volume275
Issue number43
DOIs
StatePublished - 27 Oct 2000

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