FIGURE 5.

The role of Bro1 for membrane lesion repair. (A) Growth profile on galactose-containing medium of a strain deleted for the ESCRT-III accessory factor BRO1 transformed with an empty vector (ev-αGFP) or constructs allowing for expression of αAβ₄₂wt or αAβ₄₂G37C. Cryo-EM pictures (B) and fluorescence microscopy pictures (C) of wild-type and bro1Δ cells transformed with an empty vector (ev) or expressing αAβ₄₂wt and grown for 6 h on galactose-containing medium. The indents in panel (B) zoom in on the plasma membrane and cell wall. The black arrowhead in (B) indicates a lipid droplet. Scale bars for cryo-EM pictures represent 200 nm. (D) BY4742 wild-type and a bro1Δ strains transformed with a plasmids carrying αAβ₄₂wt and additionally a plasmid allowing the expression of Kar2_(1-135)-mCherry-HDEL (ERCherry), a marker for the ER. DNA was stained with Hoechst. Cells were grown in medium allowing for gene expression for 6 h prior to microscopy. Scale bars for fluorescence pictures represent 2 μm. (E) PI staining of cells deleted for BRO1 transformed with constructs allowing for expression of αAβ₄₂wt, αAβ₄₂L34T, αAβ₄₂G37C, or αGFP after 4 or 24 h growth on galactose-containing medium. Error bars represent standard deviations of at least four independent transformants.