title: Heme Assimilation inSchizosaccharomyces pombeRequires Cell-surface-anchored Protein Shu1 and Vacuolar Transporter Abc3.
abstract: TheSchizosaccharomyces pombe shu1+gene encodes a cell-surface protein required for assimilation of exogenous heme. In this study, shaving experiments showed that Shu1 is released from membrane preparations when spheroplast lysates are incubated with phosphoinositide-specific phospholipase C (PI-PLC). Shu1 cleavability by PI-PLC and its predicted hydropathy profile strongly suggested that Shu1 is a glycosylphosphatidylinositol-anchored protein. When heme biosynthesis is selectively blocked inhem1Δ mutant cells, the heme analog zinc mesoporphyrin IX (ZnMP) first accumulates into vacuoles and then subsequently, within the cytoplasm in a rapid and Shu1-dependent manner. An HA4-taggedshu1+allele that retained wild-type function localizes to the cell surface in response to low hemin concentrations, but under high hemin concentrations, Shu1-HA4re-localizes to the vacuolar membrane. Inactivation ofabc3+, encoding a vacuolar membrane transporter, results inhem1Δabc3Δ mutant cells being unable to grow in the presence of hemin as the sole iron source. Inhem1Δabc3Δ cells, ZnMP accumulates primarily in vacuoles and does not sequentially accumulate in the cytosol. Consistent with a role for Abc3 as vacuolar hemin exporter, results with hemin-agarose pulldown assays showed that Abc3 binds to hemin. In contrast, an Abc3 mutant in which an inverted Cys-Pro motif had been replaced with Ala residues fails to bind hemin with high affinity. Taken together, these results show that Shu1 undergoes rapid hemin-induced internalization from the cell surface to the vacuolar membrane and that the transporter Abc3 participates in the mobilization of stored heme from the vacuole to the cytosol.
authors.affiliation: From the Département de Biochimie, Faculté de médecine et des sciences de la santé, Université de Sherbrooke, Sherbrooke, Quebec J1E 4K8, Canada.
authors.forename: Thierry
authors.initials: T
authors.lastname: Mourer
authors.affiliation: From the Département de Biochimie, Faculté de médecine et des sciences de la santé, Université de Sherbrooke, Sherbrooke, Quebec J1E 4K8, Canada.
authors.forename: Vincent
authors.initials: V
authors.lastname: Normant
authors.affiliation: From the Département de Biochimie, Faculté de médecine et des sciences de la santé, Université de Sherbrooke, Sherbrooke, Quebec J1E 4K8, Canada Simon.Labbe@USherbrooke.ca.
authors.forename: Simon
authors.initials: S
authors.lastname: Labbé
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D001692:Biological Transport; D006418:Heme / Q000378:metabolism*; D006427:Hemin / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D008665:Metalloporphyrins / Q000378:metabolism; D021381:Protein Transport; D012568:Schizosaccharomyces / Q000166:cytology / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000378:metabolism*
chemical_list: D018528:ATP-Binding Cassette Transporters; C548659:Abc3 protein, S pombe; D008565:Membrane Proteins; D008665:Metalloporphyrins; D029702:Schizosaccharomyces pombe Proteins; C000620284:Shu1 protein, S pombe; D006418:Heme; D006427:Hemin; C058069:zinc mesoporphyrin
keywords: ABC transporter; glycosylphosphatidylinositol (GPI anchor); heme; metal homeostasis; yeast; yeast physiology