The trafficking of subcellular cargos in eukaryotic cells crucially depends on vesicle budding, a process mediated by ARF-GEFs (ADP-ribosylation factor guanine nucleotide exchange factors). In plants, ARF-GEFs play essential roles in endocytosis, vacuolar trafficking, recycling, secretion, and polar trafficking. Moreover, they are important for plant development, mainly through controlling the polar subcellular localization of PIN-FORMED transporters of the plant hormone auxin. Here, using a chemical genetics screen in Arabidopsis thaliana, we identified Endosidin 4 (ES4), an inhibitor of eukaryotic ARF-GEFs. ES4 acts similarly to and synergistically with the established ARF-GEF inhibitor Brefeldin A and has broad effects on intracellular trafficking, including endocytosis, exocytosis, and vacuolar targeting. Additionally, Arabidopsis and yeast (Saccharomyces cerevisiae) mutants defective in ARF-GEF show altered sensitivity to ES4. ES4 interferes with the activation-based membrane association of the ARF1 GTPases, but not of their mutant variants that are activated independently of ARF-GEF activity. Biochemical approaches and docking simulations confirmed that ES4 specifically targets the SEC7 domain-containing ARF-GEFs. These observations collectively identify ES4 as a chemical tool enabling the study of ARF-GEF-mediated processes, including ARF-GEF-mediated plant development.
- MeSH
- Arabidopsis účinky léků genetika metabolismus MeSH
- brefeldin A farmakologie MeSH
- buněčná membrána účinky léků metabolismus MeSH
- chromony chemie farmakologie MeSH
- DNA vazebné proteiny genetika metabolismus MeSH
- endocytóza účinky léků MeSH
- geneticky modifikované rostliny MeSH
- membránové glykoproteiny genetika metabolismus MeSH
- membránové transportní proteiny genetika metabolismus MeSH
- mutace MeSH
- proteinové domény MeSH
- proteiny huseníčku genetika metabolismus MeSH
- Saccharomyces cerevisiae - proteiny genetika metabolismus MeSH
- Saccharomyces cerevisiae účinky léků metabolismus MeSH
- simulace molekulového dockingu MeSH
- transkripční faktory genetika metabolismus MeSH
- transport proteinů účinky léků MeSH
- výměnné faktory guaninnukleotidů chemie genetika metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- ARF1 protein, Arabidopsis MeSH Prohlížeč
- brefeldin A MeSH
- chromony MeSH
- DNA vazebné proteiny MeSH
- GNL1 protein, Arabidopsis MeSH Prohlížeč
- GNOM protein, Arabidopsis MeSH Prohlížeč
- membránové glykoproteiny MeSH
- membránové transportní proteiny MeSH
- PIN1 protein, Arabidopsis MeSH Prohlížeč
- proteiny huseníčku MeSH
- Saccharomyces cerevisiae - proteiny MeSH
- SEC12 protein, S cerevisiae MeSH Prohlížeč
- transkripční faktory MeSH
- výměnné faktory guaninnukleotidů MeSH
