Fig. 2. Identification of the insertional mutation in the REMI mutants and generation of sodC^– cells. (A) remi56 displayed an insertion at a locus encoding a superoxide dismutase (SOD) domain-containing protein, SodC. SodC encodes an N-terminal signal peptide with the C-terminal GPI-anchoring sequence (omega domain), and a partial and a full Cu/Zn-superoxide dismutase (SOD) domain. (B) sodC^– cells were generated by homologous recombination from wild-type cells, and confirmed by genomic PCR. (C) sodC^– cells showed increased basal membrane localization of GFP-PHcrac compared with wild-type cells. (D) Western blot analysis of the cytosolic and membrane fractions of the wild-type and sodC^– cells showed more GFP-PHcrac localization at the membrane in sodC than in wild-type cells. Ras proteins are shown as a loading control for membrane fractions.