Quantitative Phosphoproteomics of cipk3/9/23/26 Mutant and Wild Type in Arabidopsis thaliana

CBL-interacting protein kinases 3/9/23/26 (CIPK3/9/23/26) are central regulation components of magnesium ion homeostasis. CBL2/3 interacts with CIPK3/9/23/26, which phosphorylates their downstream targets, suggesting that protein phosphorylation is a key factor influencing the maintenance of cellular magnesium homeostasis in higher plants. The cipk3/9/23/26 quadruple mutant is very sensitive to high levels of magnesium. In this study, TMT quantitative phosphoproteomics were used to compare the global variations in phosphoproteins in wild type and cipk3/9/23/26 quadruple mutant seedlings of Arabidopsis thaliana, and 12,506 phosphorylation modification sites on 4537 proteins were identified, of which 773 phosphorylated proteins exhibited significant variations at the phosphorylation level under magnesium sensitivity. Subsequently, we used bioinformatics methods to systematically annotate and analyze the data. Certain transporters and signaling components that could be associated with magnesium sensitivity, such as ATP-binding cassette transporters and mitogen-activated protein kinases, were identified. The results of this study further our understanding of the molecular mechanisms of CIPK3/9/23/26 in mediating magnesium homeostasis.