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Proteome-wide cellular thermal shift assay reveals unexpected cross-talk between brassinosteroid and auxin signaling

Qing Lu (UGent) , Yonghong Zhang (UGent) , Joakim Hellner, Caterina Giannini, Xiangyu Xu (UGent) , Jarne Pauwels (UGent) , Qian Ma (UGent) , Wim Dejonghe (UGent) , Huibin Han, Brigitte Van De Cotte (UGent) , et al.
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Abstract
Despite the growing interest in using chemical genetics in plant research, small molecule target identification remains a major challenge. The cellular thermal shift assay coupled with highresolution mass spectrometry (CETSA MS) that monitors changes in the thermal stability of proteins caused by their interactions with small molecules, other proteins, or posttranslational modifications, allows the discovery of drug targets or the study of protein-metabolite and protein-protein interactions mainly in mammalian cells. To showcase the applicability of this method in plants, we applied CETSA MS to intact Arabidopsis thaliana cells and identified the thermal proteome of the plant-specific glycogen synthase kinase 3 (GSK3) inhibitor, bikinin. A comparison between the thermal and the phosphoproteomes of bikinin revealed the auxin efflux carrier PIN-FORMED1 (PIN1) as a substrate of the Arabidopsis GSK3s that negatively regulate the brassinosteroid signaling. We established that PIN1 phosphorylation by the GSK3s is essential for maintaining its intracellular polarity that is required for auxin-mediated regulation of vascular patterning in the leaf, thus revealing cross-talk between brassinosteroid and auxin signaling.
Keywords
auxin, brassinosteroids, cellular thermal shift assay, chemical genetics, DRUG TARGET ENGAGEMENT, CHEMICAL INHIBITION, GSK3-LIKE KINASES, GENE-EXPRESSION, ARABIDOPSIS, BIN2, PHOSPHORYLATION, TRANSDUCTION, INTEGRATION, STABILITY

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Citation

Please use this url to cite or link to this publication:

MLA
Lu, Qing, et al. “Proteome-Wide Cellular Thermal Shift Assay Reveals Unexpected Cross-Talk between Brassinosteroid and Auxin Signaling.” PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, vol. 119, no. 11, 2022, doi:10.1073/pnas.2118220119.
APA
Lu, Q., Zhang, Y., Hellner, J., Giannini, C., Xu, X., Pauwels, J., … Russinova, E. (2022). Proteome-wide cellular thermal shift assay reveals unexpected cross-talk between brassinosteroid and auxin signaling. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 119(11). https://doi.org/10.1073/pnas.2118220119
Chicago author-date
Lu, Qing, Yonghong Zhang, Joakim Hellner, Caterina Giannini, Xiangyu Xu, Jarne Pauwels, Qian Ma, et al. 2022. “Proteome-Wide Cellular Thermal Shift Assay Reveals Unexpected Cross-Talk between Brassinosteroid and Auxin Signaling.” PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 119 (11). https://doi.org/10.1073/pnas.2118220119.
Chicago author-date (all authors)
Lu, Qing, Yonghong Zhang, Joakim Hellner, Caterina Giannini, Xiangyu Xu, Jarne Pauwels, Qian Ma, Wim Dejonghe, Huibin Han, Brigitte Van De Cotte, Francis Impens, Kris Gevaert, Ive De Smet, Jiří Friml, Daniel Martinez Molina, and Eugenia Russinova. 2022. “Proteome-Wide Cellular Thermal Shift Assay Reveals Unexpected Cross-Talk between Brassinosteroid and Auxin Signaling.” PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 119 (11). doi:10.1073/pnas.2118220119.
Vancouver
1.
Lu Q, Zhang Y, Hellner J, Giannini C, Xu X, Pauwels J, et al. Proteome-wide cellular thermal shift assay reveals unexpected cross-talk between brassinosteroid and auxin signaling. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA. 2022;119(11).
IEEE
[1]
Q. Lu et al., “Proteome-wide cellular thermal shift assay reveals unexpected cross-talk between brassinosteroid and auxin signaling,” PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, vol. 119, no. 11, 2022.
@article{8745667,
  abstract     = {{Despite the growing interest in using chemical genetics in plant research, small molecule target identification remains a major challenge. The cellular thermal shift assay coupled with highresolution mass spectrometry (CETSA MS) that monitors changes in the thermal stability of proteins caused by their interactions with small molecules, other proteins, or posttranslational modifications, allows the discovery of drug targets or the study of protein-metabolite and protein-protein interactions mainly in mammalian cells. To showcase the applicability of this method in plants, we applied CETSA MS to intact Arabidopsis thaliana cells and identified the thermal proteome of the plant-specific glycogen synthase kinase 3 (GSK3) inhibitor, bikinin. A comparison between the thermal and the phosphoproteomes of bikinin revealed the auxin efflux carrier PIN-FORMED1 (PIN1) as a substrate of the Arabidopsis GSK3s that negatively regulate the brassinosteroid signaling. We established that PIN1 phosphorylation by the GSK3s is essential for maintaining its intracellular polarity that is required for auxin-mediated regulation of vascular patterning in the leaf, thus revealing cross-talk between brassinosteroid and auxin signaling.}},
  articleno    = {{e2118220119}},
  author       = {{Lu, Qing and Zhang, Yonghong and Hellner, Joakim and Giannini, Caterina and Xu, Xiangyu and Pauwels, Jarne and Ma, Qian and Dejonghe, Wim and Han, Huibin and Van De Cotte, Brigitte and Impens, Francis and Gevaert, Kris and De Smet, Ive and Friml, Jiří and Molina, Daniel Martinez and Russinova, Eugenia}},
  issn         = {{0027-8424}},
  journal      = {{PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA}},
  keywords     = {{auxin,brassinosteroids,cellular thermal shift assay,chemical genetics,DRUG TARGET ENGAGEMENT,CHEMICAL INHIBITION,GSK3-LIKE KINASES,GENE-EXPRESSION,ARABIDOPSIS,BIN2,PHOSPHORYLATION,TRANSDUCTION,INTEGRATION,STABILITY}},
  language     = {{eng}},
  number       = {{11}},
  pages        = {{9}},
  title        = {{Proteome-wide cellular thermal shift assay reveals unexpected cross-talk between brassinosteroid and auxin signaling}},
  url          = {{http://dx.doi.org/10.1073/pnas.2118220119}},
  volume       = {{119}},
  year         = {{2022}},
}

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