The central metabolic regulator SnRK1 controls plant growth and survival upon activation by energy depletion, but detailed molecular insight into its regulation and downstream targets is limited. Here we used phosphoproteomics to infer the sucrose-dependent processes targeted upon starvation by kinases as SnRK1, corroborating the relation of SnRK1 with metabolic enzymes and transcriptional regulators, while also pointing to SnRK1 control of intracellular trafficking. Next, we integrated affinity purification, proximity labelling and crosslinking mass spectrometry to map the protein interaction landscape, composition and structure of the SnRK1 heterotrimer, providing insight in its plant-specific regulation. At the intersection of this multi-dimensional interactome, we discovered a strong association of SnRK1 with class II T6P synthase (TPS)-like proteins. Biochemical and cellular assays show that TPS-like proteins function as negative regulators of SnRK1. Next to stable interactions with the TPS-like proteins, similar intricate connections were found with known regulators, suggesting that plants utilize an extended kinase complex to fine-tune SnRK1 activity for optimal responses to metabolic stress.

Ghent University Department of Plant Biotechnology and Bioinformatics Ghent Belgium

Institute of Experimental Botany Czech Academy of Sciences Prague Czech Republic

KU Leuven Plant Institute LPI Heverlee Leuven Belgium

Laboratory for Molecular Plant Biology Biology Department KU Leuven Heverlee Leuven Belgium

The Key Laboratory of Plant Development and Environmental Adaptation Biology Ministry of Education School of Life Sciences Shandong University Qingdao China

VIB Center for Plant Systems Biology Ghent Belgium

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Biomolecular condensation orchestrates clathrin-mediated endocytosis in plants

Recent insights into metabolic and signalling events of directional root growth regulation and its implications for sustainable crop production systems