FIGURE 6.

Functional expression of H165C, H165M, and C161H/H165C HMA6 mutants in yeast. A, yeast strains expressing HMA6 and HMA6 mutants were grown on selective media supplemented with copper at the indicated concentrations. For each expression condition, two independent transformants were spotted at dilutions 1 and 1/10, as 2-μl drops. Dilution 1 corresponds to an optical density of 1.8 at 600 nm. EV, empty vector. B, MIC of copper observed for each yeast strain expressing the different forms of HMA6 (the asterisks correspond to values obtained from Fig. 4). Mutants leading to the same yeast sensitivity to copper as HMA6 are indicated in bold. C, expression of HMA6 and HMA6 mutants (H165C, H165M, and C161H/H165C) in yeast membranes was revealed by Western blotting using a Strep-tactin HRP conjugate. The asterisk indicates HMA6, HMA6-H165C, HMA6-H165M, and HMA6-C161H/H165C.