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Phase separation-based visualization of protein-­protein interactions and kinase activities in plants

Alaeddine Safi (UGent) , Wouter Smagghe (UGent) , Amanda Gonçalves (UGent) , Qing Wang, Ke Xu, Ana Fernandez Salina (UGent) , Benjamin Cappe, Franck Riquet (UGent) , Evelien Mylle (UGent) , Dominique Eeckhout (UGent) , et al.
(2023) PLANT CELL. 35(9). p.3280-3302
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Abstract
Protein activities depend heavily on protein complex formation and dynamic post-translational modifications, such as phosphorylation. The dynamic nature of protein complex formation and post-translational modifications is notoriously difficult to monitor in planta at cellular resolution, often requiring extensive optimization. Here, we generated and exploited the SYnthetic Multivalency in PLants (SYMPL)-vector set to assay protein-protein interactions (PPIs) (SPPIER - Separation of Phases-based Protein Interaction Reporter) and kinase activities (SPARK - Separation of Phases-based Activity Reporter of Kinase) in planta, based on phase separation. This technology enabled easy detection of inducible, binary and ternary protein-protein interactions among cytoplasmic and nuclear proteins in plant cells via a robust image-based readout. Moreover, we applied the SYMPL toolbox to develop an in vivo reporter for SNF1-related kinase 1 (SnRK1) activity, allowing us to visualize tissue-specific, dynamic SnRK1 activity in stable transgenic Arabidopsis (Arabidopsis thaliana) plants. The SYMPL cloning toolbox provides a means to explore PPIs, phosphorylation, and other post-translational modifications with unprecedented ease and sensitivity.
Keywords
Cell Biology, Plant Science

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Citation

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MLA
Safi, Alaeddine, et al. “Phase Separation-Based Visualization of Protein-­protein Interactions and Kinase Activities in Plants.” PLANT CELL, vol. 35, no. 9, 2023, pp. 3280–302, doi:10.1093/plcell/koad188.
APA
Safi, A., Smagghe, W., Gonçalves, A., Wang, Q., Xu, K., Fernandez Salina, A., … Vanneste, S. (2023). Phase separation-based visualization of protein-­protein interactions and kinase activities in plants. PLANT CELL, 35(9), 3280–3302. https://doi.org/10.1093/plcell/koad188
Chicago author-date
Safi, Alaeddine, Wouter Smagghe, Amanda Gonçalves, Qing Wang, Ke Xu, Ana Fernandez Salina, Benjamin Cappe, et al. 2023. “Phase Separation-Based Visualization of Protein-­protein Interactions and Kinase Activities in Plants.” PLANT CELL 35 (9): 3280–3302. https://doi.org/10.1093/plcell/koad188.
Chicago author-date (all authors)
Safi, Alaeddine, Wouter Smagghe, Amanda Gonçalves, Qing Wang, Ke Xu, Ana Fernandez Salina, Benjamin Cappe, Franck Riquet, Evelien Mylle, Dominique Eeckhout, Nancy De Winne, Eveline Van De Slijke, Freya Persyn, Geert Persiau, Daniël Van Damme, Danny Geelen, Geert De Jaeger, Tom Beeckman, Jelle Van Leene, and Steffen Vanneste. 2023. “Phase Separation-Based Visualization of Protein-­protein Interactions and Kinase Activities in Plants.” PLANT CELL 35 (9): 3280–3302. doi:10.1093/plcell/koad188.
Vancouver
1.
Safi A, Smagghe W, Gonçalves A, Wang Q, Xu K, Fernandez Salina A, et al. Phase separation-based visualization of protein-­protein interactions and kinase activities in plants. PLANT CELL. 2023;35(9):3280–302.
IEEE
[1]
A. Safi et al., “Phase separation-based visualization of protein-­protein interactions and kinase activities in plants,” PLANT CELL, vol. 35, no. 9, pp. 3280–3302, 2023.
@article{01H5A1X0B800G5NDRB1DJXGKDD,
  abstract     = {{Protein activities depend heavily on protein complex formation and dynamic post-translational modifications, such as phosphorylation. The dynamic nature of protein complex formation and post-translational modifications is notoriously difficult to monitor in planta at cellular resolution, often requiring extensive optimization. Here, we generated and exploited the SYnthetic Multivalency in PLants (SYMPL)-vector set to assay protein-protein interactions (PPIs) (SPPIER - Separation of Phases-based Protein Interaction Reporter) and kinase activities (SPARK - Separation of Phases-based Activity Reporter of Kinase) in planta, based on phase separation. This technology enabled easy detection of inducible, binary and ternary protein-protein interactions among cytoplasmic and nuclear proteins in plant cells via a robust image-based readout. Moreover, we applied the SYMPL toolbox to develop an in vivo reporter for SNF1-related kinase 1 (SnRK1) activity, allowing us to visualize tissue-specific, dynamic SnRK1 activity in stable transgenic Arabidopsis (Arabidopsis thaliana) plants. The SYMPL cloning toolbox provides a means to explore PPIs, phosphorylation, and other post-translational modifications with unprecedented ease and sensitivity.}},
  author       = {{Safi, Alaeddine and Smagghe, Wouter and Gonçalves, Amanda and Wang, Qing and Xu, Ke and Fernandez Salina, Ana and Cappe, Benjamin and Riquet, Franck and Mylle, Evelien and Eeckhout, Dominique and De Winne, Nancy and Van De Slijke, Eveline and Persyn, Freya and Persiau, Geert and Van Damme, Daniël and Geelen, Danny and De Jaeger, Geert and Beeckman, Tom and Van Leene, Jelle and Vanneste, Steffen}},
  issn         = {{1040-4651}},
  journal      = {{PLANT CELL}},
  keywords     = {{Cell Biology,Plant Science}},
  language     = {{eng}},
  number       = {{9}},
  pages        = {{3280--3302}},
  title        = {{Phase separation-based visualization of protein-­protein interactions and kinase activities in plants}},
  url          = {{http://doi.org/10.1093/plcell/koad188}},
  volume       = {{35}},
  year         = {{2023}},
}
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