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Heavy metal transport by AtHMA4 involves the N‐terminal degenerated metal binding domain and the C‐terminal His 11 stretch
Author(s) -
Verret Frédéric,
Gravot Antoine,
Auroy Pascaline,
Preveral Sandra,
Forestier Cyrille,
Vavasseur Alain,
Richaud Pierre
Publication year - 2005
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2005.01.065
Subject(s) - heterologous expression , complementation , saccharomyces cerevisiae , arabidopsis , mutant , chemistry , biochemistry , cysteine , arabidopsis thaliana , bimolecular fluorescence complementation , heterologous , fusion protein , yeast , microbiology and biotechnology , biology , recombinant dna , gene , enzyme
The Arabidopsis thaliana AtHMA4 is a P 1B ‐type ATPase that clusters with the Zn/Cd/Pb/Co subgroup. It has been previously shown, by heterologous expression and the study of AtHMA4 knockout or overexpressing lines in Arabidopsis [1–3], that AtHMA4 is implicated in zinc homeostasis and cadmium tolerance. Here, we report the study of the heterologous expression of AtHMA4 in the yeast Saccharomyces cerevisiae . AtHMA4 expression resulted in an increased tolerance to Zn, Cd and Pb and to a phenotypic complementation of hypersensitive mutants. In contrast, an increased sensitivity towards Co was observed. An AtHMA4::GFP fusion protein was observed in endocytic vesicles and at the yeast plasma membrane. Mutagenesis of the cysteine and glutamate residues from the N‐ter degenerated heavy metal binding domain impaired the function of AtHMA4. It was also the case when the C‐ter His 11 stretch was deleted, giving evidence that these amino acids are essential for the AtHMA4 binding/translocation of metals.