Transport of antimony salts by Arabidopsis thaliana protoplasts over-expressing the human multidrug resistance-associated protein 1 (MRP1/ABCC1).
ABC transporters from the multidrug resistance-associated protein (MRP) subfamily are glutathione S-conjugate pumps exhibiting a broad substrate specificity illustrated by numerous xenobiotics, such as anticancer drugs, herbicides, pesticides and heavy metals. The engineering of MRP transporters into plants might be interesting either to reduce the quantity of xenobiotics taken up by the plant in the context of "safe-food" strategies or, conversely, in the development of phytoremediation strategies in which xenobiotics are sequestered in the vacuolar compartment. In this report, we obtained Arabidopsis transgenic plants overexpressing human MRP1. In these plants, expression of MRP1 did not increase plant resistance to antimony salts (Sb(III)), a classical glutathione-conjugate substrate of MRP1. However, the transporter was fully translated in roots and shoots, and targeted to the plasma membrane. In order to investigate the functionality of MRP1 in Arabidopsis, mesophyll cell protoplasts (MCPs) were isolated from transgenic plants and transport activities were measured by using calcein or Sb(III) as substrates. Expression of MRP1 at the plasma membrane was correlated with an increase in the MCPs resistance to Sb(III) and a limitation of the metalloid content in the protoplasts due to an improvement in Sb(III) efflux. Moreover, Sb(III) transport was sensitive to classical inhibitors of the human MRP1, such as MK571 or glibenclamide. These results demonstrate that a human ABC transporter can be functionally introduced in Arabidopsis, which might be useful, with the help of stronger promoters, to reduce the accumulation of xenobiotics in plants, such as heavy metals from multi-contaminated soils.
Gayet
Landry
L
CEA Cadarache, DSV-DEVM--LEMS, UMR 6191 CNRS-CEA-Université Aix-Marseille II, 13108 St Paul lez Durance, France.
Picault
Nathalie
N
Cazalé
Anne-Claire
AC
Beyly
Audrey
A
Lucas
Philippe
P
Jacquet
Hélène
H
Suso
Henri-Pierre
HP
Vavasseur
Alain
A
Peltier
Gilles
G
Forestier
Cyrille
C
D000965:Antimony / Q000737:chemistry* / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology; D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter / Q000235:genetics; D006801:Humans; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified; D014176:Protein Biosynthesis / Q000235:genetics; D011523:Protoplasts / Q000187:drug effects* / Q000378:metabolism*; D018749:RNA, Plant / Q000235:genetics; D012492:Salts / Q000737:chemistry*
D000965:Antimony; D000970:Antineoplastic Agents; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D018749:RNA, Plant; D012492:Salts; C421109:multidrug resistance-associated protein 1