{"database":"iProX","file_versions":[],"scores":null,"additional":{"omics_type":["Proteomics"],"submitter":["Shuhua Yang"],"species":["Arabidopsis Thaliana"],"full_dataset_link":["http://www.iprox.org/page/project.html?id=IPX0002607000"],"submitter_email":["yangshuhua@cau.edu.cn"],"submitter_affiliation":["China Agricultural University"],"sample_protocol":[""],"repository":["iProX"],"data_protocol":[""],"pubmed_abstract":["Protein phosphorylation and dephosphorylation have been reported to play important roles in plant cold responses. In addition, phospho-regulatory feedback is a conserved mechanism for biological processes and stress responses in animals and plants. However, it is less well known that a regulatory feedback loop is formed by the protein kinase and the protein phosphatase in plant responses to cold stress. Here, we report that OPEN STOMATA 1 (OST1) and PROTEIN PHOSPHATASE 2C G GROUP 1 (PP2CG1) reciprocally regulate the activity during the cold stress response. The interaction of PP2CG1 and OST1 is inhibited by cold stress, which results in the release of OST1 at the cytoplasm and nucleus from suppression by PP2CG1. Interestingly, cold-activated OST1 phosphorylates PP2CG1 to suppress its phosphatase activity, thereby amplifying cold signaling in plants. Mutations of PP2CG1 and its homolog PP2CG2 enhance freezing tolerance, whereas overexpression of PP2CG1 decreases freezing tolerance. Moreover, PP2CG1 negatively regulates protein levels of C-REPEAT BINDING FACTORs (CBFs) under cold stress. Our results uncover a phosphor/dephosphor-regulatory feedback loop mediated by PP2CG1 phosphatase and OST1 protein kinase in plant cold responses."],"pubmed_title":["Reciprocal regulation between the negative regulator PP2CG1 phosphatase and the positive regulator OST1 kinase confers cold response in Arabidopsis."],"pubmed_authors":["Lv Jian J, Liu Jingyan J, Ming Yuhang Y, Shi Yiting Y, Song Chunpeng C, Gong Zhizhong Z, Yang Shuhua S, Ding Yanglin Y"],"additional_accession":[]},"is_claimable":false,"name":"Cold-induced phosphorylational sites analysis of protein kinase OST1","description":"For the analysis of phosphorylation sites of OST1, 8 g pSuper:OST1-Myc plants materials, treated with 4oC for 0 or 2 h, were used for protein extraction and immunoprecipitated with anti-Myc agarose. Then OST1-Myc proteins were eluted with 0.1 M glycine solution (pH 2.8) and mixed with 1 M Tris solution to ensure the pH of OST1-Myc is 7.5. After the concentration of PP2CG1 proteins with 30K-ultrafiltration tubes, phosphatase assay was performed: OST1-Myc was treated with GST-PP2CG1 for 0 or 60 min under 30oC. Then, the samples were digested with trypsin using FASP method. The collected solution containing digested peptide were lyophilized for phospho-peptides enrichment, which was performed using TiO2-tips (Thermo Scientific, #A32992) according to the manufacture’s protocol. The phosphor-peptides were resuspended in 0.1% FA for nano LC-MS analysis.","dates":{"publication":"Mon Nov 16 00:00:00 GMT 2020"},"accession":"PXD022539","cross_references":{"TAXONOMY":["3702"],"pubmed":["33871153"]}}