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

Kiril Mishev, Qing Lu, Bram Denoo, François Peurois, Wim Dejonghe, Jan Hullaert, Riet De Rycke, Sjef Boeren, Marine Bretou, Steven De Munck, Isha Sharma, Kaija Goodman, Kamila Kalinowska, Veronique Storme, Le Son Long Nguyen, Andrzej Drozdzecki, Sara Martins, Wim Nerinckx, Dominique Audenaert, Grégory VertAnnemieke Madder, Marisa S. Otegui, Erika Isono, Savvas N. Savvides, Wim Annaert, Sacco de Vries, Jacqueline Cherfils, Johan Winne, Eugenia Russinova*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

7 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)2573-2593
JournalThe Plant Cell
Volume30
Issue number10
DOIs
Publication statusPublished - 1 Oct 2018

Fingerprint Dive into the research topics of 'Nonselective Chemical Inhibition of Sec7 Domain-Containing ARF GTPase Exchange Factors'. Together they form a unique fingerprint.

  • Cite this

    Mishev, K., Lu, Q., Denoo, B., Peurois, F., Dejonghe, W., Hullaert, J., De Rycke, R., Boeren, S., Bretou, M., De Munck, S., Sharma, I., Goodman, K., Kalinowska, K., Storme, V., Nguyen, L. S. L., Drozdzecki, A., Martins, S., Nerinckx, W., Audenaert, D., ... Russinova, E. (2018). Nonselective Chemical Inhibition of Sec7 Domain-Containing ARF GTPase Exchange Factors. The Plant Cell, 30(10), 2573-2593. https://doi.org/10.1105/tpc.18.00145