title: A common highly conserved cadmium detoxification mechanism from bacteria to humans: heavy metal tolerance conferred by the ATP-binding cassette (ABC) transporter SpHMT1 requires glutathione but not metal-chelating phytochelatin peptides.
abstract: Cadmium poses a significant threat to human health due to its toxicity. In mammals and in bakers' yeast, cadmium is detoxified by ATP-binding cassette transporters after conjugation to glutathione. In fission yeast, phytochelatins constitute the co-substrate with cadmium for the transporter SpHMT1. In plants, a detoxification mechanism similar to the one in fission yeast is supposed, but the molecular nature of the transporter is still lacking. To investigate further the relationship between SpHMT1 and its co-substrate, we overexpressed the transporter in a Schizosaccharomyces pombe strain deleted for the phytochelatin synthase gene and heterologously in Saccharomyces cerevisiae and in Escherichia coli. In all organisms, overexpression of SpHMT1 conferred a markedly enhanced tolerance to cadmium but not to Sb(III), AgNO(3), As(III), As(V), CuSO(4), or HgCl(2). Abolishment of the catalytic activity by expression of SpHMT1(K623M) mutant suppressed the cadmium tolerance phenotype independently of the presence of phytochelatins. Depletion of the glutathione pool inhibited the SpHMT1 activity but not that of AtHMA4, a P-type ATPase, indicating that GSH is necessary for the SpHMT1-mediated cadmium resistance. In E. coli, SpHMT1 was targeted to the periplasmic membrane and led to an increased amount of cadmium in the periplasm. These results demonstrate that SpHMT1 confers cadmium tolerance in the absence of phytochelatins but depending on the presence of GSH and ATP. Our results challenge the dogma of the two separate cadmium detoxification pathways and demonstrate that a common highly conserved mechanism has been selected during the evolution from bacteria to humans.
authors.affiliation: Commissariat à l'Energie Atomique (CEA) Cadarache, Service de Biologie Végétale et de Microbiologie Environnementales, Laboratoire des Echanges Membranaires et Signalisation, the CNRS, UMR Biologie Végétale et de Microbiologie Environnementales.
authors.forename: Sandra
authors.initials: S
authors.lastname: Prévéral
authors.forename: Landry
authors.initials: L
authors.lastname: Gayet
authors.forename: Cristina
authors.initials: C
authors.lastname: Moldes
authors.forename: Jonathan
authors.initials: J
authors.lastname: Hoffmann
authors.forename: Sandra
authors.initials: S
authors.lastname: Mounicou
authors.forename: Antoine
authors.initials: A
authors.lastname: Gruet
authors.forename: Florie
authors.initials: F
authors.lastname: Reynaud
authors.forename: Ryszard
authors.initials: R
authors.lastname: Lobinski
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Verbavatz
authors.forename: Alain
authors.initials: A
authors.lastname: Vavasseur
authors.forename: Cyrille
authors.initials: C
authors.lastname: Forestier
mesh_terms: D018528:ATP-Binding Cassette Transporters; D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000235:genetics / Q000378:metabolism*; D019943:Amino Acid Substitution; D019881:Aminoacyltransferases / Q000235:genetics / Q000378:metabolism; D000818:Animals; D017360:Arabidopsis; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D002104:Cadmium / Q000494:pharmacology*; D002614:Chelating Agents; D025141:Drug Resistance, Fungal / Q000502:physiology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D055786:Gene Knockout Techniques; D005978:Glutathione / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D020125:Mutation, Missense; D054811:Phytochelatins; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D012568:Schizosaccharomyces / Q000235:genetics / Q000378:metabolism*
chemical_list: D018528:ATP-Binding Cassette Transporters; D029681:Arabidopsis Proteins; D002614:Chelating Agents; D011994:Recombinant Proteins; C489670:hmt1 protein, S pombe; D002104:Cadmium; D000255:Adenosine Triphosphate; D054811:Phytochelatins; D019881:Aminoacyltransferases; C093784:glutathione gamma-glutamylcysteinyltransferase; D000251:Adenosine Triphosphatases; C477864:HMA4 protein, Arabidopsis; D005978:Glutathione