High level of Hrq1-7KR results in cell lethality.

(A) Hrq1-7KR protein levels are similar to WT in basal conditions. The indicative strains were grown overnight in 2% galactose, subsequently TCA was performed and western was run to determined protein level. (B) Stabilization or overexpression of Hrq1 does not lead to increased cisplatin sensitivity. The indicated yeast strains were five-fold serial diluted onto SC medium containing DMSO and/or SC medium containing the indicated amount of cisplatin. The plates were photographed after 2 days of incubation at 30°C in the dark. Plates that were used with Hrq1-OE (GAL-HRQ1, galactose inducible/dextrose repressible promoter) strains were made with galactose instead of glucose medium. (C) Overexpression of the Hrq1-7KR mutant results in cell lethality. The indicated yeast strains were grown overnight in SC medium containing raffinose. Subsequently they were five-fold serial diluted onto galactose SC plates. The plates were photographed after 2 days of incubation at 30°C in the dark. (D) pCMV-RECQL4-KA promotes similar overexpression compared to pCMV-RECQL4. Protein extracts from cells 3 days post-transfection. Endogenous RECQL4 was detected using α-RECQL4, GAPDH served as a control. (TIF)