The ABC transporter AtPDR8 is a cadmium extrusion pump conferring heavy metal resistance.
Cadmium (Cd) and lead (Pb) are widespread pollutants that are toxic to plant growth. The expression of AtPDR8 was upregulated in cadmium- or lead-treated Arabidopsis thaliana. To test whether AtPDR8 is involved in heavy metal resistance, we examined transgenic Arabidopsis that over-expressed AtPDR8 and RNAi plants that exhibited a severely reduced AtPDR8 transcript level, as well as T-DNA insertion mutants of this ABC transporter. AtPDR8-over-expressing plants were more resistant to Cd(2+) or Pb(2+) than the wild-type and had lower Cd contents. In contrast, AtPDR8 RNAi transgenic plants and T-DNA insertion lines were more sensitive to Cd(2+) or Pb(2+) compared to wild-type plants and had higher Cd contents. The GFP-AtPDR8 protein was targeted to the plasma membrane, and GUS activity was present in most cells but strongest in the root hair and epidermal cells. Cd extrusion was higher in the AtPDR8-over-expressing plants in a flux assay using isolated protoplasts and radioactive (109)Cd, and was lower in the RNAi transgenic plants than in the wild-type. Together, these data strongly support a role for AtPDR8 as an efflux pump of Cd(2+) or Cd conjugates at the plasma membrane of Arabidopsis cells. As AtPDR8 has been suggested to be involved in the pathogen response and in the transport of chemicals that mediate pathogen resistance, this ABC protein is likely to transport a very broad range of substrates.
Kim
Do-Young
DY
POSTECH-UZH Cooperative Laboratory, Division of Molecular Life Sciences, Pohang University of Science and Technology, Pohang 790-784, Korea.
Bovet
Lucien
L
Maeshima
Masayoshi
M
Martinoia
Enrico
E
Lee
Youngsook
Y
D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000502:physiology*; D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000502:physiology*; D001692:Biological Transport / Q000187:drug effects; D015152:Blotting, Northern; D015153:Blotting, Western; D002104:Cadmium / Q000378:metabolism / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D004351:Drug Resistance; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007854:Lead / Q000378:metabolism / Q000494:pharmacology; D019216:Metals, Heavy / Q000378:metabolism* / Q000494:pharmacology; D018517:Plant Roots / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D011523:Protoplasts / Q000187:drug effects / Q000378:metabolism; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction
D018528:ATP-Binding Cassette Transporters; D029681:Arabidopsis Proteins; D019216:Metals, Heavy; D002104:Cadmium; D049452:Green Fluorescent Proteins; D007854:Lead