Figure 3
From: Distinct functional relevance of dynamic GTPase cysteine methylation in fission yeast
Mam4 mediates proper plasma membrane tethering of palmitoylated and farnesylated GTPases lacking polybasic motifs. (a) Deconvolved images of cells from control and mam4∆ strains expressing genomic unprenylated GFP-Rho2-HA-RitC fusions grown in YES medium and observed by fluorescence microscopy. Representative fluorescence intensity plots (as arbitrary fluorescence units) were generated from line scans across the cell width (dotted white lines). (b) Images of cells from control and mam4∆ strains expressing genomic geranylgeranylated and palmitoylated GFP-Rho2-HA-CCIIL fusions observed by fluorescence microscopy. Representative fluorescence intensity plots were obtained as described in (a). (c) Deconvolved images of mixed control (GFP-Rho2-HA-CCIIL, mCherry-Atb2) and mam4∆ (GFP-Rho2-HA-CCIIL) cells were observed by fluorescence microscopy. (d) Images of cells from control and mam4∆ strains expressing genomic polybasic, farnesylated and palmitoylated GFP-Rho2-HA-polyB fusions observed by fluorescence microscopy. Representative fluorescence intensity plots were obtained as described in (a). (e) Images of cells from control and mam4∆ strains expressing genomic farnesylated and palmitoylated GFP-Rho1-CCIIS fusions (Rho2 tail) were observed by fluorescence microscopy. Representative fluorescence intensity plots were obtained as described in (a). (f) Deconvolved images of mixed control (GFP-Rho1-CCIIS, mCherry-Atb2) and mam4∆ (GFP-Rho1-CCIIS) cells were observed by fluorescence microscopy. (G) Cell extracts from growing cultures in YES medium of strains with the indicated genotypes were resolved by SDS-PAGE and hybridized separately with anti-GFP and anti-Cdc2 (loading control) antibodies. Results representative of two independent experiments are shown.
