Fig. 4.

CgSkn7 and CgYap1 cooperatively bind to oxidative stress inducible promoters. (A) Analysis of the promoter regions for significantly enriched Yap1/Skn7 binding site pairs located in close proximity. (B) CgSkn7-CFP is constitutively localized in the nucleus. CgSkn7-CFP was visualized by fluorescence microscopy of Cgskn7Δ mutant cells transformed with pCgADH1-CgSKN7-CFP. Upon oxidative stress, GFP-CgYap1 is localized in the nucleus in Cgyap1Δskn7Δ mutant cells. Cgyap1Δskn7Δ cells transformed with pCgADH1-GFP-CgYAP1 were grown in synthetic medium. GFP-CgYap1 was visualized by fluorescence microscopy (0.4 mM hydrogen peroxide, 10 min). (C) Chromatin recruitment assay of CYap1 and CgSkn7. Cgyap1Δ, Cgskn7Δ and Cgyap1Δskn7Δ mutant strains transformed with pHA-CgYAP1 (C) or pCgSKN7-HA (D) were grown to early exponential phase (OD 0.4) and treated with 0.4 mM H₂O₂ for times indicated. Anti-HA precipitated DNA was analyzed by qPCR.