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Proteome-wide cellular thermal shift assay reveals novel crosstalk between brassinosteroid and auxin signaling

ABSTRACT: We demonstrated that bikinin changed the thermal stability of some of its target proteins as well as several known or putative interacting proteins of Arabidopsis GSK3s. By combining the thermal- and the phospho-proteomes of bikinin we uncovered the auxin carrier PIN-FORMED1 (PIN1) as a novel substrate of the Arabidopsis GSK3s. We showed that inhibition the kinase activity of GSK3s by brassinosteroids or the specific chemical inhibitor bikinin led to the loss of PIN1 polarity, revealing a novel crosstalk between brassinosteroid and auxin signaling. Hence, our study demonstrates the applicability of CETSA MS in plant intact cells for identification of small-molecule targets and for discovery of novel protein-protein interactions.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Arabidopsis Thaliana (ncbitaxon:3702)

SUBMITTER: Eugenia Russinova

PROVIDER: MSV000088545 | MassIVE | Wed Dec 08 02:35:00 GMT 2021

REPOSITORIES: MassIVE

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Project description:This is the single cell model for analysis of hormonal crosstalk in Arabidopsis described in the article: Modelling and experimental analysis of hormonal crosstalk in Arabidopsis. Liu J, Mehdi S, Topping J, Tarkowski P and Lindsey K. Mol Syst Biol. 2010 Jun 8;6:373; PmID: 20531403 , DOI: 10.1038/msb.2010.26 Abstract: An important question in plant biology is how genes influence the crosstalk between hormones to regulate growth. In this study, we model POLARIS (PLS) gene function and crosstalk between auxin, ethylene and cytokinin in Arabidopsis. Experimental evidence suggests that PLS acts on or close to the ethylene receptor ETR1, and a mathematical model describing possible PLS-ethylene pathway interactions is developed, and used to make quantitative predictions about PLS-hormone interactions. Modelling correctly predicts experimental results for the effect of the pls gene mutation on endogenous cytokinin concentration. Modelling also reveals a role for PLS in auxin biosynthesis in addition to a role in auxin transport. The model reproduces available mutants, and with new experimental data provides new insights into how PLS regulates auxin concentration, by controlling the relative contribution of auxin transport and biosynthesis and by integrating auxin, ethylene and cytokinin signalling. Modelling further reveals that a bell-shaped dose-response relationship between endogenous auxin and root length is established via PLS. This combined modelling and experimental analysis provides new insights into the integration of hormonal signals in plants. This model was originally created using Copasi and taken from the supplementary materials of the MSB article. It uses equation 5 for the auxin biosynthesis and was altered to also contain the reactions for ACC, IAA and cytokinine import. Different from the supplementary material, the parameters for the auxin synthesis, v2, are set to k2c = 0.01 uM and k2=0.2 uM_per_sec and for the WT PLS transcription k6=0.3 . To obtain the model described in the first table of the supplementary materials, set k2c=k2=0 and k6=0.9 . For the pls and PLSox mutants, k6 should be set to 0 and 0.45, respectively. This model originates from BioModels Database: A Database of Annotated Published Models. It is copyright (c) 2005-2010 The BioModels Team. For more information see the terms of use . To cite BioModels Database, please use Le Novère N., Bornstein B., Broicher A., Courtot M., Donizelli M., Dharuri H., Li L., Sauro H., Schilstra M., Shapiro B., Snoep J.L., Hucka M. (2006) BioModels Database: A Free, Centralized Database of Curated, Published, Quantitative Kinetic Models of Biochemical and Cellular Systems Nucleic Acids Res., 34: D689-D691.

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Proteome-wide cellular thermal shift assay reveals novel crosstalk between brassinosteroid and auxin signaling

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