Figure 7. A conserved residue in the Sit1 SITD compromises ferrichrome utilization and survival in macrophages.

A The Sit1^Y575A strain is compromised in ferrichrome utilization under Fe deficiency. C. glabrata strains were grown to mid-logarithmic phase and 10-fold serial dilutions spotted onto rich synthetic media and Fe-deficient media supplemented or not with 10 µM ferrichrome. B Sit1^Y575A localizes to the plasma membrane. In vivo fluorescence of wild type Sit1mCherry and Sit1^Y575AmCherry strains cultured under Fe deficiency. C The Sit1^Y575A strain is compromised the ability to use siderophores to survive macrophage killing. Activated J774A.1 cells were infected with wild type, sit1Δ or sit1Δ::SIT1^Y575A C. glabrata strains and CFU quantitated. Results are shown as survival relative to wild type C. glabrata grown under Fe deficiency represented by a dashed line. Each experiment included 6 replicates per experimental sample. Results are shown as mean survival compared to the wild type grown in the absence of ferrichrome supplementation and are representative of at least 3 experiments. Bars represent the standard error.