CadA of Mesorhizobium metallidurans isolated from a zinc-rich mining soil is a PIB-2-type ATPase involved in cadmium and zinc resistance
2014
Maynaud, Géraldine | Brunel, Brigitte | Yashiro, Erika | Mergeay, Max | Cleyet-Marel, Jean Claude | Le Quéré, Antoine | Laboratoire des symbioses tropicales et méditerranéennes (UMR LSTM) ; Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Université Montpellier 1 (UM1)-Institut de Recherche pour le Développement (IRD)-Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université de Montpellier (UM)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro) | Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro) | Molecular and Cellular Biology, Unit Microbiology ; Centre d'Etude de l'Energie Nucléaire (SCK-CEN) | Agence Nationale de la Recherche Programme “Contaminants, Ecosystèmes et Santé” (CES 2008-12, APTITUDE).
Mesorhizobium metallidurans STM 2683T is a nitrogen-fixing bacterium that nodulates Anthyllis vulneraria in mine tailings highly contaminated in zinc, lead and cadmium. To study the mechanisms whereby this bacterium copes with metals, we functionally screened a cosmid genomic library of M. metallidurans for zinc or cadmium tolerance. A cosmid clone harbored a gene encoding PIB-type ATPase homologous to CadA that leads to cadmium and zinc resistance in Escherichia coli. The CadA protein structure presents one duplication of the two N-terminal metal binding domains (i.e. a heavy metal-associated domain followed by a histidine-rich domain) which allows specific binding to zinc and cadmium cations. A cadA-deleted strain of M. metallidurans failed to grow at high zinc concentrations (2 mM) and its growth was delayed at lower zinc concentrations. Expression studies using a transcriptional fusion of cadA promoter to gfp showed that cadA is specifically induced in a dose-dependent manner by zinc and cadmium in M. metallidurans in vitro conditions and into A. vulneraria nodules after Zn stress. Metal induction sensitivity was increased in the strain where cadA gene was deleted.This study identified cadA as a first mesorhizobial resistance determinant involved in detoxification of cadmium and zinc and which confers upon M. metallidurans greater capacity for coping with high zinc concentrations. This improves the knowledge of this bacterium for potential use as a symbiotic inoculant of Anthyllis in phytostabilization strategies of metal-rich sites.
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