Figure S2. Δubc8 shows increased precursor levels.
(A) Wild-type and Δubc8 cells were precultured in a lactate medium, and expression of cytosolic DHFR (−) and b2(1-167)Δ19-DHFR (+) was induced by the addition of 0.5% galactose for 4 h. Samples were taken, and cell lysates were analyzed by Western blot. Precursor (p) and mature (m) forms of Mdj1 were detected. Sod1 served as a loading control. The arrows point at the precursor form of Mdj1. (B) Rpn4-mediated stress response was measured using a reporter expressing the yellow fluorescent protein under the control of a proteasome-associated control element (Boos et al, 2019). Proteasome subunits are expressed under the control of proteasome-associated control elements, and their expression is induced upon mitoprotein-induced stress conditions. Significance testing was performed using a t test (n.s., not significant). (C) UBC8 gene was deleted in two different genetic backgrounds, and growth on glucose and the non-fermentable carbon source glycerol was tested. (D) A mitochondria-targeted GFP protein was expressed in the cells indicated (Dimmer et al, 2002). Cells were grown in galactose medium before the morphology of the mitochondrial structure was visualized by fluorescence microscopy. The Δmdm12 mutant was used as a control for a strain with altered morphology (Dimmer et al, 2002). Scale bar = 5 μm.
Source data are available for this figure.
