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

Alaeddine Safi; Wouter Smagghe; Amanda Gonçalves; Qing Wang; Ke Xu; Ana Ibis Fernandez; Benjamin Cappe; Franck B. 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; Steffen Vanneste

doi:10.1093/plcell/koad188

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

Alaeddine Safi, Wouter Smagghe, Amanda Gonçalves, Qing Wang, Ke Xu, Ana Ibis Fernandez, Benjamin Cappe, Franck B. 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, Steffen Vanneste

Research output: Contribution to journal › Article › peer-review

10 Scopus citations

Abstract

Protein activities depend heavily on protein complex formation and dynamic posttranslational modifications, such as phosphorylation. The dynamic nature of protein complex formation and posttranslational 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) (separation of phases-based protein interaction reporter) and kinase activities (separation of phases-based activity reporter of kinase) in planta, based on phase separation. This technology enabled easy detection of inducible, binary and ternary PPIs 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 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 posttranslational modifications with unprecedented ease and sensitivity.

Original language	English
Pages (from-to)	3280-3302
Number of pages	23
Journal	Plant Cell
Volume	35
Issue number	9
DOIs	https://doi.org/10.1093/plcell/koad188
State	Published - 1 Sep 2023
Externally published	Yes

ASJC Scopus subject areas

Plant Science

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10.1093/plcell/koad188

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Safi, A., Smagghe, W., Gonçalves, A., Wang, Q., Xu, K., Fernandez, A. I., Cappe, B., Riquet, F. B., Mylle, E., Eeckhout, D., De Winne, N., Van De Slijke, E., Persyn, F., Persiau, G., Van Damme, D., Geelen, D., De Jaeger, G., Beeckman, T., Van Leene, J., & 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

@article{631d0666639e4fd0af95d83a664051e7,

title = "Phase separation-based visualization of protein-protein interactions and kinase activities in plants",

abstract = "Protein activities depend heavily on protein complex formation and dynamic posttranslational modifications, such as phosphorylation. The dynamic nature of protein complex formation and posttranslational 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) (separation of phases-based protein interaction reporter) and kinase activities (separation of phases-based activity reporter of kinase) in planta, based on phase separation. This technology enabled easy detection of inducible, binary and ternary PPIs 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 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 posttranslational modifications with unprecedented ease and sensitivity.",

author = "Alaeddine Safi and Wouter Smagghe and Amanda Gon{\c c}alves and Qing Wang and Ke Xu and Fernandez, \{Ana Ibis\} and Benjamin Cappe and Riquet, \{Franck B.\} and Evelien Mylle and Dominique Eeckhout and \{De Winne\}, Nancy and \{Van De Slijke\}, Eveline and Freya Persyn and Geert Persiau and \{Van Damme\}, Dani{\"e}l and Danny Geelen and \{De Jaeger\}, Geert and Tom Beeckman and \{Van Leene\}, Jelle and Steffen Vanneste",

year = "2023",

month = sep,

day = "1",

doi = "10.1093/plcell/koad188",

language = "English",

volume = "35",

pages = "3280--3302",

journal = "Plant Cell",

issn = "1040-4651",

publisher = "American Society of Plant Biologists",

number = "9",

}

Safi, A, Smagghe, W, Gonçalves, A, Wang, Q, Xu, K, Fernandez, AI, Cappe, B, Riquet, FB, Mylle, E, Eeckhout, D, De Winne, N, Van De Slijke, E, Persyn, F, Persiau, G, Van Damme, D, Geelen, D, De Jaeger, G, Beeckman, T, Van Leene, J & Vanneste, S 2023, 'Phase separation-based visualization of protein-protein interactions and kinase activities in plants', Plant Cell, vol. 35, no. 9, pp. 3280-3302. https://doi.org/10.1093/plcell/koad188

TY - JOUR

T1 - Phase separation-based visualization of protein-protein interactions and kinase activities in plants

AU - Safi, Alaeddine

AU - Smagghe, Wouter

AU - Gonçalves, Amanda

AU - Wang, Qing

AU - Xu, Ke

AU - Fernandez, Ana Ibis

AU - Cappe, Benjamin

AU - Riquet, Franck B.

AU - Mylle, Evelien

AU - Eeckhout, Dominique

AU - De Winne, Nancy

AU - Van De Slijke, Eveline

AU - Persyn, Freya

AU - Persiau, Geert

AU - Van Damme, Daniël

AU - Geelen, Danny

AU - De Jaeger, Geert

AU - Beeckman, Tom

AU - Van Leene, Jelle

AU - Vanneste, Steffen

PY - 2023/9/1

Y1 - 2023/9/1

N2 - Protein activities depend heavily on protein complex formation and dynamic posttranslational modifications, such as phosphorylation. The dynamic nature of protein complex formation and posttranslational 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) (separation of phases-based protein interaction reporter) and kinase activities (separation of phases-based activity reporter of kinase) in planta, based on phase separation. This technology enabled easy detection of inducible, binary and ternary PPIs 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 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 posttranslational modifications with unprecedented ease and sensitivity.

AB - Protein activities depend heavily on protein complex formation and dynamic posttranslational modifications, such as phosphorylation. The dynamic nature of protein complex formation and posttranslational 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) (separation of phases-based protein interaction reporter) and kinase activities (separation of phases-based activity reporter of kinase) in planta, based on phase separation. This technology enabled easy detection of inducible, binary and ternary PPIs 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 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 posttranslational modifications with unprecedented ease and sensitivity.

UR - https://www.scopus.com/pages/publications/85169715114

U2 - 10.1093/plcell/koad188

DO - 10.1093/plcell/koad188

M3 - Article

C2 - 37378595

AN - SCOPUS:85169715114

SN - 1040-4651

VL - 35

SP - 3280

EP - 3302

JO - Plant Cell

JF - Plant Cell

IS - 9

ER -

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

Abstract

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