A Systems Biology Approach Reveals the Role of a Novel Methyltransferase in Response to Chemical Stress and Lipid Homeostasis
Figure 6
Crg1 is important for actin patch formation during cantharidin treatment.
(A) Diagram showing how PIP species are involved in Pkc1-dependent changes in actin cytoskeleton. (B) Crg1 is important for actin patch integrity during cantharidin stress. Wt, crg1Δ/Δ mutant and CRG1-overexpressing crg1Δ/Δ cells were grown to mid-exponential phase at 30°C in YPD in the presence of cantharidin (250 µM) or DMSO for 1 hour. Cells fixed with formaldehyde were stained for actin with FITC-phalloidin and visualized by fluorescence microscopy. Bar, 5 µm. The number of actin patches per cell in each sample was quantified. Values are the mean of three independent replicates (n = 270–1000), error bars are the standard deviation; * P-value <0.025, ** P-value <0.0002 (Student's t-test). (C) Cantharidin induces CRG1 transcription via the Cell Wall Integrity (CWI) pathway. Cells grown to mid-exponential phase were treated with cantharidin (250 µM) in YPD for indicated time. Total RNA was extracted, cDNA was prepared and analyzed by qRT-PCR. CRG1 transcript levels were normalized to ACT1. The simplified diagram of CWI pathway is shown. Slt2 is a kinase, Rlm1 is a transcriptional activator governed by the CWI pathway. (D) A preliminary model describing Crg1-cantharidin interaction. See text for details.