Novel MAP kinase substrates identified by solid-phase phosphorylation screening in Arabidopsis thaliana

Title:	Novel MAP kinase substrates identified by solid-phase phosphorylation screening in Arabidopsis thaliana
Authors:	Park, H.C. Nguyen, X.C. Bahk, S., (...) Bohnert, H.J. Chung, W.S.
Keywords:	Arabidopsis;MAP kinase;Phosphorylation;Solid-phase screening;Substrates
Issue Date:	2016
Publisher:	Springer Tokyo
Citation:	Scopus
Abstract:	Phosphorylation of substrate proteins by mitogen-activated protein kinases (MPKs) determines the specific cellular responses elicited by a particular extracellular stimulus. However, downstream targets of plant MPKs remain poorly characterized. In this study, 29 putative substrates of AtMPK3, AtMPK4 and AtMPK6 were identified by solid-phase phosphorylation screening of a λ phage expression library constructed from combined mRNAs from salt-treated, pathogen-treated and mechanically wounded Arabidopsis seedlings. To test the efficiency of this screening, we performed in vitro kinase assay with 10 recombinant fusion proteins. All proteins were phosphorylated by AtMPK3, AtMPK4 and AtMPK6, indicating the efficiency of this screening procedure. To confirm phosphorylation of isolated substrates by plant MPKs, we performed in-gel kinase assays. All test substrates were strongly phosphorylated by wounding or H2O2-activated AtMPK3 and AtMPK6. Three substrates, encoded by genes At2g41430, At2g41900, and At3g16770, were strongly phosphorylated, suggesting a function as AtMPK substrates. The type of screening provides a powerful way for identifying potential substrates of MAP kinases responsive to biotic and abiotic stresses.
Description:	Plant Biotechnology Reports Volume 10, Issue 6, 1 November 2016, Pages 415-423
URI:	http://link.springer.com/article/10.1007%2Fs11816-016-0412-9 http://repository.vnu.edu.vn/handle/VNU_123/33274
ISSN:	18635466
Appears in Collections:	Bài báo của ĐHQGHN trong Scopus