Fig. 2: Glycerol synthesis is required for proper Arl1 and Imh1 localization.
From: Glycerol mediates crosstalk between metabolism and trafficking through the golgin Imh1
a, Glycerol synthesis pathway in S. cerevisiae. The glycolytic metabolite DHAP is converted to G3P by GPD1 and GPD2 and glycerol is then produced by the phosphatases GPP1 and GPP2 (ref. 26). b, Arl1 and Imh1 were mislocalized in gpd1∆ and gpp1/2∆ cells. Deletion of the upstream Arl1 regulator Arl3 and the Arl1 guanine-nucleotide exchange factor Syt1 resulted in mislocalization of Arl1 and Imh1. mCH–Imh1 or Arl1–mRFP was coexpressed with Sec7–GFP in the indicated strains. Live cells were observed in midlog phase. Scale bars, 5 μm. In a,b, the ratio of colocalization between Imh1 or Arl1 and Golgi marker Sec7 was quantified (N = 3, n = 50). The data are presented as the mean ± s.d. of three independent experiments. Statistical analysis was conducted using a one-way ANOVA with Dunnett’s post hoc multiple-comparison test. The WT strain was used as a reference. c,d, Gpd1 and Gpp1/2 regulate the localization of Imh1. c, The enzymatic activity of Gpd1 is required for proper Imh1 localization. mCH–Imh1, Sec7–GFP and Gpd1 constructs were coexpressed in cells (Gpd1-K245A, enzymatically inactive Gpd1 mutant). d, Gpp1/2 are required for Imh1 proper localization. mCH–Imh1, Sec7–GFP, and Gpp2 constructs were coexpressed in cells. Live cells were observed in the midlog phase. The numbers of mCH–Imh1 puncta were determined with ImageJ Fiji software (N = 3, n = 50). Data are presented as the mean ± s.d. of three independent experiments. Statistical analysis was conducted using a one-way ANOVA with Dunnett’s post hoc multiple-comparison test. The WT strain was used as a reference. Scale bars, 5 μm.
