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Nonselective Chemical Inhibition of Sec7 Domain-Containing ARF GTPase Exchange Factors

Kiril Mishev (UGent) , Qing Lu (UGent) , Bram Denoo (UGent) , Francois Peurois, Wim Dejonghe (UGent) , Jan Hullaert (UGent) , Riet De Rycke (UGent) , Sjef Boeren, Marine Bretou, Steven De Munck (UGent) , et al.
(2018) PLANT CELL. 30(10). p.2573-2593
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Abstract
Small GTP-binding proteins from the ADP-ribosylation factor (ARF) family are important regulators of vesicle formation and cellular trafficking in all eukaryotes. ARF activation is accomplished by a protein family of guanine nucleotide exchange factors (GEFs) that contain a conserved catalytic Sec7 domain. Here, we identified and characterized Secdin, a small-molecule inhibitor of Arabidopsis thaliana ARF-GEFs. Secdin application caused aberrant retention of plasma membrane (PM) proteins in late endosomal compartments, enhanced vacuolar degradation, impaired protein recycling, and delayed secretion and endocytosis. Combined treatments with Secdin and the known ARF-GEF inhibitor Brefeldin A (BFA) prevented the BFA-induced PM stabilization of the ARF-GEF GNOM, impaired its translocation from the Golgi to the trans-Golgi network/early endosomes, and led to the formation of hybrid endomembrane compartments reminiscent of those in ARF-GEF-deficient mutants. Drug affinity-responsive target stability assays revealed that Secdin, unlike BFA, targeted all examined Arabidopsis ARF-GEFs, but that the interaction was probably not mediated by the Sec7 domain because Secdin did not interfere with the Sec7 domain-mediated ARF activation. These results show that Secdin and BFA affect their protein targets through distinct mechanisms, in turn showing the usefulness of Secdin in studies in which ARF-GEF-dependent endomembrane transport cannot be manipulated with BFA.
Keywords
GOLGI NETWORK/EARLY ENDOSOME, RECEPTOR KINASE BRI1, ARABIDOPSIS-THALIANA, PLANT-CELLS, PLASMA-MEMBRANE, ENDOMEMBRANE, TRAFFICKING, PROTEIN COMPLEXES, SMALL MOLECULES, WIDE ANALYSIS, BREFELDIN-A

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Chicago
Mishev, Kiril, Qing Lu, Bram Denoo, Francois Peurois, Wim Dejonghe, Jan Hullaert, Riet De Rycke, et al. 2018. “Nonselective Chemical Inhibition of Sec7 Domain-Containing ARF GTPase Exchange Factors.” Plant Cell 30 (10): 2573–2593.
APA
Mishev, K., Lu, Q., Denoo, B., Peurois, F., Dejonghe, W., Hullaert, J., De Rycke, R., et al. (2018). Nonselective Chemical Inhibition of Sec7 Domain-Containing ARF GTPase Exchange Factors. PLANT CELL, 30(10), 2573–2593.
Vancouver
1.
Mishev K, Lu Q, Denoo B, Peurois F, Dejonghe W, Hullaert J, et al. Nonselective Chemical Inhibition of Sec7 Domain-Containing ARF GTPase Exchange Factors. PLANT CELL. 2018;30(10):2573–93.
MLA
Mishev, Kiril et al. “Nonselective Chemical Inhibition of Sec7 Domain-Containing ARF GTPase Exchange Factors.” PLANT CELL 30.10 (2018): 2573–2593. Print.
@article{8586099,
  abstract     = {Small GTP-binding proteins from the ADP-ribosylation factor (ARF) family are important regulators of vesicle formation and cellular trafficking in all eukaryotes. ARF activation is accomplished by a protein family of guanine nucleotide exchange factors (GEFs) that contain a conserved catalytic Sec7 domain. Here, we identified and characterized Secdin, a small-molecule inhibitor of Arabidopsis thaliana ARF-GEFs. Secdin application caused aberrant retention of plasma membrane (PM) proteins in late endosomal compartments, enhanced vacuolar degradation, impaired protein recycling, and delayed secretion and endocytosis. Combined treatments with Secdin and the known ARF-GEF inhibitor Brefeldin A (BFA) prevented the BFA-induced PM stabilization of the ARF-GEF GNOM, impaired its translocation from the Golgi to the trans-Golgi network/early endosomes, and led to the formation of hybrid endomembrane compartments reminiscent of those in ARF-GEF-deficient mutants. Drug affinity-responsive target stability assays revealed that Secdin, unlike BFA, targeted all examined Arabidopsis ARF-GEFs, but that the interaction was probably not mediated by the Sec7 domain because Secdin did not interfere with the Sec7 domain-mediated ARF activation. These results show that Secdin and BFA affect their protein targets through distinct mechanisms, in turn showing the usefulness of Secdin in studies in which ARF-GEF-dependent endomembrane transport cannot be manipulated with BFA.},
  author       = {Mishev, Kiril and Lu, Qing and Denoo, Bram and Peurois, Francois and Dejonghe, Wim and Hullaert, Jan and De Rycke, Riet and Boeren, Sjef and Bretou, Marine and De Munck, Steven and Sharma, Isha and Goodman, Kaija and Kalinowska, Kamila and Storme, Veronique and Nguyen, Long and Drozdzecki, Andrzej and Martins, Sara and Nerinckx, Wim and Audenaert, Dominique and Vert, Gregory and Madder, Annemieke and Otegui, Marisa S. and Isono, Erika and Savvides, Savvas and Annaert, Wim and De Vries, Sacco and Cherfils, Jacqueline and Winne, Johan and Russinova, Eugenia},
  issn         = {1040-4651},
  journal      = {PLANT CELL},
  language     = {eng},
  number       = {10},
  pages        = {2573--2593},
  title        = {Nonselective Chemical Inhibition of Sec7 Domain-Containing ARF GTPase Exchange Factors},
  url          = {http://dx.doi.org/10.1105/tpc.18.00145},
  volume       = {30},
  year         = {2018},
}

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