Functional characterization of AtATM1, AtATM2, and AtATM3, a subfamily of Arabidopsis half-molecule ATP-binding cassette transporters implicated in iron homeostasis.
The functional capabilities of one of the smallest subfamilies of ATP-binding cassette transporters from Arabidopsis thaliana, the AtATMs, are described. Designated AtATM1, AtAATM2, and AtATM3, these half-molecule ABC proteins are homologous to the yeast mitochondrial membrane protein ATM1 (ScATM1), which is clearly implicated in the export of mitochondrially synthesized iron/sulfur clusters. Yeast ATM1-deficient (atm1) mutants grow very slowly (have a petite phenotype), are respiration-deficient, accumulate toxic levels of iron in their mitochondria, and show enhanced compensatory high affinity iron uptake. Of the three Arabidopsis ATMs, AtATM3 bears the closest functional resemblance to ScATM1. Heterologously expressed AtATM3 is not only able to complement the yeast atm1 petite phenotype but is also able to suppress the constitutively high capacity for high affinity iron uptake associated with loss of the chromosomal copy of ScATM1, abrogate intra-mitochondrial iron hyperaccumulation, and restore mitochondrial respiratory function and cytochrome c levels. By comparison, AtATM1 only weakly suppresses the atm1 phenotype, and AtATM2 exerts little or no suppressive action but instead is toxic when expressed in this system. The differences between AtATM3 and AtATM1 are maintained after exchanging their target peptides, and these proteins as well as AtATM2 colocalize with the mitochondrial fluor MitoTracker Red when expressed in yeast as GFP fusions. Although its toxicity when heterologously expressed in yeast, except when fused with GFP, precludes the functional analysis of native AtATM2, a common function, mitochondrial export of Fe/S clusters or their precursors for the assembly of cytosolic Fe/S proteins, is inferred for AtATM3 and AtATM1.
Chen
Sixue
S
Plant Science Institute, Department of Biology, Carolyn Hoff Lynch Biology Laboratory, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Sánchez-Fernández
Rocío
R
Lyver
Elise R
ER
Dancis
Andrew
A
Rea
Philip A
PA
D018528:ATP-Binding Cassette Transporters / Q000737:chemistry / Q000502:physiology*; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000502:physiology*; D003001:Cloning, Molecular; D045304:Cytochromes c / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D007501:Iron / Q000737:chemistry / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D008957:Models, Genetic; D009154:Mutation; D010641:Phenotype; D010802:Phylogeny; D018517:Plant Roots / Q000378:metabolism; D021381:Protein Transport
C522743:ATM1 protein, Arabidopsis; C522744:ATM2 protein, Arabidopsis; C510573:ATM3 protein, Arabidopsis; D018528:ATP-Binding Cassette Transporters; D029681:Arabidopsis Proteins; D045304:Cytochromes c; D007501:Iron