A drug‐resistant mutation in plant target of rapamycin validates the specificity of ATP ‐competitive TOR inhibitors in vivo - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Plant Journal Année : 2023

A drug‐resistant mutation in plant target of rapamycin validates the specificity of ATP ‐competitive TOR inhibitors in vivo

Résumé

Kinases are major components of cellular signaling pathways, regulating key cellular activities through phosphorylation. Kinase inhibitors are efficient tools for studying kinase targets and functions, however assessing their kinase specificity in vivo is essential. The identification of resistant kinase mutants has been proposed to be the most convincing approach to achieve this goal. Here, we address this issue in plants via a pharmacogenetic screen for mutants resistant to the ATP‐competitive TOR inhibitor AZD‐8055. The eukaryotic TOR (Target of Rapamycin) kinase is emerging as a major hub controlling growth responses in plants largely thanks to the use of ATP‐competitive inhibitors. We identified a dominant mutation in the DFG motif of the Arabidopsis TOR kinase domain that leads to very strong resistance to AZD‐8055. This resistance was characterized by measuring root growth, photosystem II (PSII) activity in leaves and phosphorylation of YAK1 (Yet Another Kinase 1) and RPS6 (Ribosomal protein S6), a direct and an indirect target of TOR respectively. Using other ATP‐competitive TOR inhibitors, we also show that the dominant mutation is particularly efficient for resistance to drugs structurally related to AZD‐8055. Altogether, this proof‐of‐concept study demonstrates that a pharmacogenetic screen in Arabidopsis can be used to successfully identify the target of a kinase inhibitor in vivo and therefore to demonstrate inhibitor specificity. Thanks to the conservation of kinase families in eukaryotes, and the possibility of creating amino acid substitutions by genome editing, this work has great potential for extending studies on the evolution of signaling pathways in eukaryotes.
Fichier principal
Vignette du fichier
Perdoux et al 2023 The Plant Journal.pdf (2.5 Mo) Télécharger le fichier
Origine Publication financée par une institution
Licence

Dates et versions

hal-04312618 , version 1 (28-11-2023)

Licence

Identifiants

Citer

Romain Perdoux, Adam Barrada, Manal Boulaiz, Camille Garau, Clément Belbachir, et al.. A drug‐resistant mutation in plant target of rapamycin validates the specificity of ATP ‐competitive TOR inhibitors in vivo. Plant Journal, 2023, 117, pp.13441355. ⟨10.1111/tpj.16564⟩. ⟨hal-04312618⟩
19 Consultations
16 Téléchargements

Altmetric

Partager

Gmail Mastodon Facebook X LinkedIn More