Fig. 7: yArf1 regulates LD-associated functions and β-oxidation.
From: Arf1 coordinates fatty acid metabolism and mitochondrial homeostasis
a, Schematic of TAG synthesis and breakdown on LD. Key enzymes involved in TAG synthesis (Dga1), hydrolysis (Tgl4), and FA activation (Faa1) used in our co-IP experiments are shown in green. b,d,e, Co-IP of yArf1–GFP and yArf1-11–GFP with Dga1-6xHA (b), Tgl4-6xHA (d) and Faa1-6xHA (e). Strains were grown at 23 °C or shifted to 37 °C. c, Co-localization of yArf1–GFP and yArf1-11–GFP with the diacylglycerol acyltransferase Dga1 tagged with 3xmCherry grown at 23 °C or shifted to 37 °C for 30 min. Arrows indicate sites of co-localization or juxtaposition between the yArf1/yArf1-11 and Dga1 on the ER or on LD. Scale bars, 5 µm. f, Peroxisome biogenesis followed by microscopy using the peroxisomal marker Pex3 fused to mCherry in the yARF1 and yarf1-11 strains grown at 23 °C or shifted to 37 °C (left). Quantification of peroxisomes per cell in each strain and condition (right). Mean and standard deviation are shown; yARF1 + Pex3–mCherry 23 °C = 1,496 cells, yARF1 + Pex3–mCherry 37 °C = 1,070 cells, yarf1-11 + Pex3–mCherry 23 °C = 1,025 cells, yarf1-11 + Pex3–mCherry 23 °C = 970 cells from n = 3 biological replicates. Scale bars, 5 µm. g, Schematic of TAG mobilization to synthesize acetyl-CoA by peroxisomal β-oxidation in yeast. Relevant proteins monitored in h are shown. Vlc-FA, very-long-chain FAs. h, Immunoblot analysis of all β-oxidation proteins, both acyl-CoA transporters and the Vlc-FA transporter genomically fused to 6xHA in the yARF1 and yarf1-11 strains grown at 23 °C or shifted to 37 °C. Pgk1 was used as loading control. i, Relative fold changes in protein levels from immunodetections done in h. Mean and standard deviation are shown from n = 3 biological replicates. Source numerical data and unprocessed blots are available in source data. See also Extended Data Fig. 5.
