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. 2001 Apr;12(4):1161–1175. doi: 10.1091/mbc.12.4.1161

Figure 2.

Figure 2

(A–D) Complementation of cdc3-124 ts strain at 36°C by expression of mutant profilins from the weakest nmt-1 promoter in pRep81 plasmid in the absence of thiamine. (A) Actin binding mutants. (B) Actin binding mutants, wild-type profilin (WT), mutant profilin (E42K) for cdc3-124 strain, and structure stability control mutant C89S. (C) Poly-l-proline binding mutants. (D) Mutants with profound loss of poly-l-proline binding Y5D, actin binding K81E, and nucleotide exchange activity Y79R, mutant with increased nucleotide exchange activity K81F, ts mutant (E42K), wild-type (WT) profilin, vector control as well as cdc3-124 strain. (E) Immunoblots to measure the expression levels of mutant profilins in cdc3-124 ts strain shifted to 36°C for 4 h. Approximately 25 μg of protein was loaded in each lane. Table 1 lists the quantitation of these results. No profilin was detected in the cdc3-124 strain without transformation or when transformed with mutant E42K.