NET4 and RabG3 link actin to the tonoplast and facilitate cytoskeletal remodelling during stomatal immunity

Timothy J. Hawkins, Michaela Kopischke, Patrick J. Duckney, Katarzyna Rybak, David A. Mentlak, Johan T. M. Kroon, Mai Thu Bui, A. Christine Richardson, Mary Casey, Agnieszka Alexander, Geert Jaeger, Monika Kalde, Ian Moore, Yasin Dagdas, Patrick J. Hussey () and Silke Robatzek ()
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Timothy J. Hawkins: University of Durham
Michaela Kopischke: Norwich Research Park
Patrick J. Duckney: University of Durham
Katarzyna Rybak: LMU Munich Biocenter
David A. Mentlak: University of Durham
Johan T. M. Kroon: University of Durham
Mai Thu Bui: Austrian Academy of Sciences, Vienna BioCenter
A. Christine Richardson: University of Durham
Mary Casey: Norwich Research Park
Agnieszka Alexander: Norwich Research Park
Geert Jaeger: VIB-University Ghent, Center for Plant System Biology
Monika Kalde: Department of Plant Sciences, University of Oxford
Ian Moore: Department of Plant Sciences, University of Oxford
Yasin Dagdas: Austrian Academy of Sciences, Vienna BioCenter
Patrick J. Hussey: University of Durham
Silke Robatzek: Norwich Research Park

Nature Communications, 2023, vol. 14, issue 1, 1-19

Abstract: Abstract Members of the NETWORKED (NET) family are involved in actin-membrane interactions. Here we show that two members of the NET family, NET4A and NET4B, are essential for normal guard cell actin reorganization, which is a process critical for stomatal closure in plant immunity. NET4 proteins interact with F-actin and with members of the Rab7 GTPase RABG3 family through two distinct domains, allowing for simultaneous localization to actin filaments and the tonoplast. NET4 proteins interact with GTP-bound, active RABG3 members, suggesting their function being downstream effectors. We also show that RABG3b is critical for stomatal closure induced by microbial patterns. Taken together, we conclude that the actin cytoskeletal remodelling during stomatal closure involves a molecular link between actin filaments and the tonoplast, which is mediated by the NET4-RABG3b interaction. We propose that stomatal closure to microbial patterns involves the coordinated action of immune-triggered osmotic changes and actin cytoskeletal remodelling likely driving compact vacuolar morphologies.

Date: 2023
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DOI: 10.1038/s41467-023-41337-z

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