abc3+ encodes an iron-regulated vacuolar ABC-type transporter in Schizosaccharomyces pombe.
Studies have shown the fundamental contribution of the yeast vacuole as a site for storage and detoxification of metals. Whereas the transmembrane proteins responsible for iron transport into and out of the vacuole have been identified in Saccharomyces cerevisiae, less information is available concerning the mobilization of vacuolar iron stores in Schizosaccharomyces pombe. In this study, we report the identification of a gene designated abc3(+) that encodes a protein which exhibits sequence homology with the ABCC subfamily of ATP-binding cassette transporters. The transcription of abc3(+) is induced by low concentrations of iron but repressed by high levels of iron. The iron-mediated repression of abc3(+) required a functional fep1(+) gene. Chromatin immunoprecipitation assays showed that Fep1 associates with the abc3(+) promoter in vivo, in an iron-dependent manner. Microscopic analyses revealed that a functional Abc3-green fluorescent protein localizes to the membrane vacuole when iron levels were low. Abc3 was required for growth in low-iron medium in the absence of the transport system mediated by Fio1 and Fip1. abc3Delta cells exhibited increased levels of expression of the frp1(+)-encoded ferric reductase, suggesting a loss of Fep1 repression and, consequently, the activation of Fep1-regulated genes. When abc3(+) was expressed using the nmt1(+) promoter system, its induction led to a reduced transcriptional activity of the frp1(+) gene. Because S. pombe does not possess vacuolar membrane-localized orthologs to S. cerevisiae Fth1, Fet5, and Smf3, our findings suggested that Abc3 may be responsible for mobilizing stored iron from the vacuole to the cytosol in response to iron deficiency.
Pouliot
Benoît
B
Département de Biochimie, Université de Sherbrooke, Sherbrooke, Quebec J1H 5N4, Canada.
Jbel
Mehdi
M
Mercier
Alexandre
A
Labbé
Simon
S
D018528:ATP-Binding Cassette Transporters / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D050980:GATA Transcription Factors / Q000235:genetics / Q000378:metabolism; D007501:Iron / Q000378:metabolism*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012568:Schizosaccharomyces / Q000235:genetics / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics* / Q000378:metabolism*; D014617:Vacuoles / Q000378:metabolism*
D018528:ATP-Binding Cassette Transporters; C548659:Abc3 protein, S pombe; C459859:Fep1 protein, S pombe; D050980:GATA Transcription Factors; D011993:Recombinant Fusion Proteins; D029702:Schizosaccharomyces pombe Proteins; D007501:Iron