title: A yeast manganese transporter related to the macrophage protein involved in conferring resistance to mycobacteria.
abstract: A novel Saccharomyces cerevisiae mutant, unable to grow in the presence of 12.5 mM EGTA, was isolated by replica plating. The phenotype of the mutant is caused by a single amino acid change (Gly149 to Arg) in the essential yeast gene CDC1. The mutant could be suppressed by overexpression of the SMF1 gene, which was isolated as an extragenic high-copy suppressor. The SMF1 gene codes for a highly hydrophobic protein and its deletion renders the yeast cells sensitive to low manganese concentration. In accordance with this observation, the smf1 null mutant exhibits reduced Mn2+ uptake at micromolar concentrations. Using a specific antibody, we demonstrated that Smf1p is located in the yeast plasma membrane. These results suggest that Smf1p is involved in high-affinity Mn2+ uptake. This assumption was also tested by overexpressing the SMF1 gene in the temperature-sensitive mutant of the mitochondrial processing peptidase (MAS1). SMF1 overexpression as well as addition of 1 mM Mn2+ to the growth medium complemented this mutation. This also suggests that in vivo Mas1p is a manganese-dependent peptidase. The yeast Smf1p resembles a protein from Drosophila and mammalian macrophages. The latter was implicated in conferring resistance to mycobacteria. A connection between Mn2+ transport and resistance or sensitivity to mycobacteria is discussed.
authors.affiliation: Roche Institute of Molecular Biology, Nutley, NJ 07110, USA.
authors.forename: F
authors.initials: F
authors.lastname: Supek
authors.forename: L
authors.initials: L
authors.lastname: Supekova
authors.forename: H
authors.initials: H
authors.lastname: Nelson
authors.forename: N
authors.initials: N
authors.lastname: Nelson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001693:Biological Transport, Active; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D008264:Macrophages / Q000378:metabolism* / Q000382:microbiology*; D008345:Manganese / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000472:pathogenicity*; D010641:Phenotype; D017354:Point Mutation; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D000096122:Mitochondrial Processing Peptidase
chemical_list: D002352:Carrier Proteins; D005656:Fungal Proteins; D008345:Manganese; D008565:Membrane Proteins; D008666:Metalloendopeptidases; D000096122:Mitochondrial Processing Peptidase