Extended Data Fig. 1: Overexpression of Mge1 leads to upregulation of mitoCPR target genes.
From: A direct role for a mitochondrial targeting sequence in signalling stress
(a) Live-cell fluorescence imaging of cells expressing PTEF2-GFP-PDR3 and the nuclear marker HTB1-mCherry under control (empty vector) and impaired mitochondrial protein import (PSD1 overexpression) conditions. Zoom-in images of representative cells are shown on the right. Scale bars, 5 μm. OE, overexpression. (b) CIS1 and PSD1 mRNA level in cells used in (a) and main Fig. 1a. n = 3 biological replicates; Two-tailed Student’s t-test. OE, overexpression. (c) PSD1 mRNA level in cells used for ChIP analysis (Fig. 1b). One-way ANOVA followed by Tukey’s test. (d) CIS1 mRNA levels in control cells (empty vector, n = 3 biological repeats) and cells overexpressing the indicated mitochondrial genes (n = 3 biological repeats). Overexpression was induced by the addition of galactose for 4 h. One-way ANOVA followed by Dunnett’s test; **** P ≤ 0.0001. OE, overexpression. (e) mRNA levels of the indicated overexpressed genes (n = 3 biological repeats) compared to empty vector control (n = 2 biological repeats) following 4-h galactose induction. Two-tailed Student’s t-test. EV, empty vector, OE, overexpression (f) Same as (d). The indicated overexpressed mitochondrial genes were tagged with 2XV5 at the N-terminus. (g) Same analysis as (e). (h) Immunoblot of overexpressed Mge1-HA-ProtA and 2V5-Mge1-HA-ProtA following 4-h galactose induction in wild-type and PDR3 deleted cells. OE, overexpression. (i) mRNA levels of YGR035C, PDR5, PDR15, GRE2 and BDH2 in cells overexpressing MGE1 and 2V5-MGE1 for 4 h. n = 3 biological replicates; One-way ANOVA followed by Dunnett’s test. OE, overexpression. (b-g, i) Data represent mean +/− SD; ns, not significant.
