The iron transporter Fth1p forms a complex with the Fet5 iron oxidase and resides on the vacuolar membrane.
Iron transport across the plasma membrane appears to be a unidirectional process whereby iron uptake is essentially irreversible. One of the major sequestration sites for iron is the vacuole that stores a variety of metals, either as a mechanism to detoxify the cell or as a reservoir of metal to enable the cell to grow when challenged by a low iron environment. Exactly how the vacuole contributes to the overall iron metabolism of the cell is unclear because mutations that affect vacuolar function also perturb the assembly of the plasma membrane high affinity transport system composed of a copper-containing iron oxidase, Fet3p, and an Fe(3+)-specific iron transporter, Ftr1p. Here, we characterize the iron transporter homologue Fth1p, which is similar to the high affinity plasma membrane iron transporter Ftr1p. We found that Fth1p was localized to the vacuolar surface and, like other proteins that function on the vacuole, did not undergo Pep4-dependent degradation. Co-immunoprecipitation experiments showed that Fth1p also associates with the Fet3p oxidase homologue, Fet5p; and disruption of the FET5 gene results in the accumulation of Fth1p in the endoplasmic reticulum. We also found that loss of this protein complex leads to elevated transcriptional activity of the FET3 gene and compromises the ability of the cell to switch from fermentative metabolism to respiratory metabolism. Because the Fet5 protein is oriented such that the oxidase domain of Fet5p is lumenal, this complex may be responsible for mobilizing intravacuolar stores of iron.
Urbanowski
J L
JL
Department of Physiology, University of Iowa, Iowa City, Iowa 52242, USA.
Piper
R C
RC
D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002570:Ceruloplasmin / Q000378:metabolism*; D017931:DNA Primers; D049452:Green Fluorescent Proteins; D006706:Homeostasis; D007425:Intracellular Membranes / Q000378:metabolism*; D007501:Iron / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D014617:Vacuoles / Q000378:metabolism*
D002352:Carrier Proteins; D017931:DNA Primers; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D049452:Green Fluorescent Proteins; D007501:Iron; D002570:Ceruloplasmin; C085513:FET3 protein, S cerevisiae; C500094:FET5 protein, S cerevisiae