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. 2020 Jan 8;5(1):e00818-19. doi: 10.1128/mSphere.00818-19

FIG 4.

FIG 4

PUUP interferes with Hsp90 activity. (A) Comparison of the transcript profile of PUUP with previously reported transcript profiles for the response to heat shock and the Hsp90 inhibitor celastrol. A diagrammatic representation of the comparison is shown for 80 genes that were induced by PUUP and also induced by either heat shock or celastrol. Colored boxes indicate upregulated genes. Red asterisks indicate genes identified in the YEASTRACT database that are induced by Hsf1. (B) Comparison of the 29 mutants identified in the genome-wide fitness profile of PUUP with previously reported mutants displaying hypersensitivity to either elevated temperature or the Hsp90 inhibitors geldanamycin and macbecin II. A diagrammatic representation of the comparison is shown, with colored boxes indicating hypersensitive mutants. Genes deleted in the mutants are listed on the left. Red asterisks indicate genes which encode proteins that interact with the yeast Hsp90 isoforms Hsp82 and Hsc82, as reported in the BioGRID database. (C) WT yeast cells containing an HSE-lacZ reporter were treated with DMSO or PUUP for 4 h, and β-Gal activity was measured. Values shown are the mean ± SD from triplicate samples. (D) WT yeast cells transformed with a glucocorticoid receptor assay system were treated with DMSO or PUUP along with 20 μM DOC or vehicle for 2 h, and β-Gal activity was measured. Values shown are the mean ± SD from triplicate samples. (E) Fivefold dilutions of parent strain BY4742 and strains containing plasmids overexpressing CDC37, HSC82, HSP82, and SLT2 were inoculated on agar plates containing DMSO or PUUP and incubated for 3 days. Two independent transformant colonies gave similar results. v-SNARE, vesicle SNAP REceptor; MAPK, mitogen-activated protein kinase; LCB, long-chain base; GPI, glycophosphatidylinositol; ER, endoplasmic reticulum; ESCRT, endosomal sorting complexes required for transport; TF, transcription factor; GEF, guanine nucleotide exchange factor; SWI/SNF, SWItch/Sucrose NonFermentable; GARP, Golgi-associated retrograde protein.