1. Pupylation-based proximity labeling reveals regulatory factors in cellulose biosynthesis in Arabidopsis.
- Author
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Zheng S, Noack LC, Khammy O, De Meyer A, Khan GA, De Winne N, Eeckhout D, Van Damme D, and Persson S
- Subjects
- Endocytosis, Cell Membrane metabolism, Staining and Labeling methods, Protein Interaction Mapping methods, Clathrin metabolism, Glucosyltransferases, Arabidopsis metabolism, Arabidopsis genetics, Cellulose biosynthesis, Cellulose metabolism, Arabidopsis Proteins metabolism, Arabidopsis Proteins genetics
- Abstract
Knowledge about how and where proteins interact provides a pillar for cell biology. Protein proximity-labeling has emerged as an important tool to detect protein interactions. Biotin-related proximity labeling approaches are by far the most commonly used but may have labeling-related drawbacks. Here, we use pupylation-based proximity labeling (PUP-IT) as a tool for protein interaction detection in plants. We show that PUP-IT readily confirmed protein interactions for several known protein complexes across different types of plant hosts and that the approach increased detection of specific interactions as compared to biotin-based proximity labeling systems. To further demonstrate the power of PUP-IT, we used the system to identify protein interactions of the protein complex that underpin cellulose synthesis in plants. Apart from known complex components, we identified the ARF-GEF BEN1 (BFA-VISUALIZED ENDOCYTIC TRAFFICKING DEFECTIVE1). We show that BEN1 contributes to cellulose synthesis by regulating both clathrin-dependent and -independent endocytosis of the cellulose synthesis protein complex from the plasma membrane. Our results highlight PUP-IT as a powerful proximity labeling system to identify protein interactions in plant cells., Competing Interests: Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)
- Published
- 2025
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