Functional importance of the βCOP appendage domain. A. Schematic illustrating the characteristics of the various Sec26p truncation constructs used for experiments. B. Plasmids coding for truncated constructs of SEC26, wild type SEC26, or vector only were transformed into the sec26Δ tester strain and spotted onto complete media (YPD) and synthetic complete media (SCD) with 5-FOA at 25, 30, 34, 37, and 40°C. C. Plasmids coding for vector alone or full length or truncated constructs under control of the copper-inducible promoter (PCUP1) were transformed into wild type yeast and replica-plated onto minimal media (SD) with and without the addition of Cu2+ (0 or 0.3 mM CuSO4). Growth was followed for three or seven days (3d and 7d respectively). The dominant negative yip1-6 construct was included as a negative control . D. Transformed cells were grown in liquid minimal media with and without the exogenous CuSO4. Cultures were grown to early log phase and adjusted to an OD600 of 0.2. CuSO4 was added at the levels indicated and cells were grown at 30°C with turbidity measured every half hour for 2 days.
DeRegis et al. BMC Cell Biology 2008 9:3 doi:10.1186/1471-2121-9-3