The Yeast Cell Fusion Protein Prm1p Requires Covalent Dimerization to Promote Membrane Fusion
Figure 5
Charged residues within a conserved hydrophobic region adjacent to Cys-120 promote Prm1p covalent dimerization and polarized localization.
(A) ClustalW alignment of the Prm1p hydrophobic region adjacent to Cys-120. (B) Anti-GFP Western blot on whole cell lysate after expression of prm1-E104L-GFP or prm1-D112L-GFP was induced with 10 µg/ml α-factor for 90 min. (C) MATa cells coexpressing prm1-D112L-GFP and prm1-D112L-myc were mated to wild-type MATα cells on YPD plates at 30°C for 3 h. Myc-tagged fusion proteins were immunoprecipitated using α-myc-Agarose. (D) Transmitted light and epifluorescence images of prm1D112LGFP in a live mating pair.