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. 2010 Aug 20;22(8):2908–2922. doi: 10.1105/tpc.110.076265

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© 2010 American Society of Plant Biologists

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Figure 5. — Induced Expression of clp1 Blocks Pheromone-Induced Conjugation Tube Formation and Cell Cycle Arrest in an rbf1-Dependent Manner.

(A) Strains JB1 (a1 Δb), UVO151 (a1 Δb; P_crg1:clp1), and UVO151Δrbf1 (a1 Δb; Δrbf1 P_crg1:clp1) were treated with synthetic a2 pheromone (2.5 μg/mL) for 6 h in MM-NA. DMSO, which was used as solvent for the a2 pheromone, served as negative control. Strains JB1 and UVO151Δrbf1 respond to pheromone treatment with the formation of conjugation tubes (arrows). By contrast, cells of strain UVO151 do not form conjugation tubes, but continue to grow by budding, indicating that the cell cycle is not arrested upon pheromone treatment in that strain. Bar = 15 μm.

(B) Pheromone-dependent gene expression in strains JB1 (a1 Δb), UVO151 (a1 Δb; P_crg1:clp1), and UVO151Δrbf1 (a1 Δb; Δrbf1; P_crg1:clp1). Cells were treated as described above, and gene expression of the a mating-type locus genes mfa1 and pra1 and of the pheromone response factors prf1 and rbf1 was determined using qRT-PCR. Shown are mean values of two technical replicates. Error bars represent the sd. Actin and eIF2b were used for normalization. The experiment was repeated three times with similar results.