Biochemical properties of vacuolar zinc transport systems of Saccharomyces cerevisiae.
The yeast vacuole plays an important role in zinc homeostasis by storing zinc for later use under deficient conditions, sequestering excess zinc for its detoxification, and buffering rapid changes in intracellular zinc levels. The mechanisms involved in vacuolar zinc sequestration are only poorly characterized. Here we describe the properties of zinc transport systems in yeast vacuolar membrane vesicles. The major zinc transport activities in these vesicles were ATP-dependent, requiring a H+ gradient generated by the V-ATPase for function. One system we identified was dependent on the ZRC1 gene, which encodes a member of the cation diffusion facilitator family of metal transporters. These data are consistent with the proposed role of Zrc1 as a vacuolar zinc transporter. Zrc1-independent activity was also observed that was not dependent on the closely related vacuolar Cot1 protein. Both Zrc1-dependent and independent activities showed a high specificity for Zn(2+) over other physiologically relevant substrates such as Ca2+, Fe2+, and Mn2+. Moreover, these systems had high affinities for zinc with apparent K(m) values in the 100-200 nm range. These results provide biochemical insight into the important role of Zrc1 and related proteins in eukaryotic zinc homeostasis.
MacDiarmid
Colin W
CW
Departments of Nutritional Sciences and Physiology, University of Missouri, Columbia, Missouri 65211, USA.
Milanick
Mark A
MA
Eide
David J
DJ
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D001692:Biological Transport; D002118:Calcium / Q000378:metabolism; D027682:Cation Transport Proteins / Q000378:metabolism; D002412:Cations; D002462:Cell Membrane / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D007477:Ions; D007501:Iron / Q000378:metabolism; D007700:Kinetics; D008345:Manganese / Q000378:metabolism; D026901:Membrane Transport Proteins; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D013997:Time Factors; D025262:Vacuolar Proton-Translocating ATPases / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism; D015032:Zinc / Q000378:metabolism*
C076223:COT1 protein, S cerevisiae; D027682:Cation Transport Proteins; D002412:Cations; D005656:Fungal Proteins; D007477:Ions; D026901:Membrane Transport Proteins; D029701:Saccharomyces cerevisiae Proteins; C088843:ZRC1 protein, S cerevisiae; D008345:Manganese; D000255:Adenosine Triphosphate; D007501:Iron; D000251:Adenosine Triphosphatases; D025262:Vacuolar Proton-Translocating ATPases; D015032:Zinc; D002118:Calcium