Figure 2.

Complementation of the CAAX protease-deficient Δrce1 Δste24 mutant by RCE1∷HA and HA∷STE24 and immunodetection of the epitope-tagged proteins. (A) Strains were tested for their mating efficiency under stringent conditions by a patch mating assay, with the SM1068 (MATα lys1) mating partner, as described (11). The growth of prototrophic diploids is indicative of mating. MATa strains tested are: SM3614, SM1058/pRS316, SM3614/pSM1093, SM3614/pSM1107, SM3614/pSM1275, SM3614/pSM1314 for a-f), respectively. (B) Blots were probed with the anti-HA antibody (lanes 1–4) or the Rce1p antibody (lanes 5–8), as described in Materials and Methods. The approximate molecular mass (kDa) of a set of protein standards is indicated for each blot. Strains used are: SM3614/pRS316, SM3614/pSM1107, SM3614/pSM1314, SM1058/pSM1097 and pSM1314, SM3613, SM1058/pRS314, SM3613/pSM1275, and SM1058/pHY01 for lanes 1–8, respectively.