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Identification of novel interactors and potential phosphorylation substrates of GsSnRK1 from wild soybean ( Glycine soja )
Author(s) -
Song Yu,
Zhang Hang,
You Hongguang,
Liu Yuanming,
Chen Chao,
Feng Xu,
Yu Xingyu,
Wu Shengyang,
Wang Libo,
Zhong Shihua,
Li Qiang,
Zhu Yanming,
Ding Xiaodong
Publication year - 2019
Publication title -
plant, cell and environment
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.646
H-Index - 200
eISSN - 1365-3040
pISSN - 0140-7791
DOI - 10.1111/pce.13217
Subject(s) - bimolecular fluorescence complementation , biology , kinase , biochemistry , abiotic stress , phosphorylation , protein subunit , complementation , transcription factor , yeast , microbiology and biotechnology , gene , phenotype
Abstract The plant sucrose nonfermenting kinase 1 (SnRK1) kinases play the central roles in the processes of energy balance, hormone perception, stress resistance, metabolism, growth, and development. However, the functions of these kinases are still elusive. In this study, we used GsSnRK1 of wild soybean as bait to perform library‐scale screens by the means of yeast two‐hybrid to identify its interacting proteins. The putative interactions were verified by yeast retransformation and β‐galactosidase assays, and the selected interactions were further confirmed in planta by bimolecular fluorescence complementation and biochemical Co‐IP assays. Protein phosphorylation analyses were carried out by phos‐tag assay and anti‐phospho‐(Ser/Thr) substrate antibodies. Finally, we obtained 24 GsSnRK1 interactors and several putative substrates that can be categorized into SnRK1 regulatory β subunit, protein modification, biotic and abiotic stress‐related, hormone perception and signalling, gene expression regulation, water and nitrogen transport, metabolism, and unknown proteins. Intriguingly, we first discovered that GsSnRK1 interacted with and phosphorylated the components of soybean nodulation and symbiotic nitrogen fixation. The interactions and potential functions of GsSnRK1 and its associated proteins were extensively discussed and analysed. This work provides plausible clues to elucidate the novel functions of SnRK1 in response to variable environmental, metabolic, and physiological requirements.