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Protein phosphorylation associated with drought priming-enhanced heat tolerance in a temperate grass species

Horticulture Research 2020年第1期7卷 309-320页

作者： Xiaxiang Zhang Lili Zhuang Yu Liu Zhimin Yang Bingru Huang College of Agro-grassland Science Nanjing Agricultural University210095 NanjingChina Department of Plant Biology Rutgers UniversityNew BrunswickNJ 08901USA

Protein phosphorylation is known to play crucial roles in plant tolerance to individual stresses,but how protein phosphorylation is associated with cross-stress tolerance,particularly drought priming-enhanced heat tolerance is largely unknown.The objectives of the present study were to identify phosphorylated proteins and phosphorylation sites that were responsive to drought priming and to determine whether drought priming-enhanced heat tolerance in temperate grass species involves changes in protein phosphorylation.Comparative analysis of phosphoproteomic profiles was performed on leaves of tall fescue(Festuca arundinacea)exposed to heat stress(38/33°C,day/night)with or without drought priming.A total of 569 differentially regulated phosphoproteins(DRPs)with 1098 phosphorylation sites were identified in response to drought priming or heat stress individually or sequentially.Most DRPs were nuclear-localized and cytosolic proteins.Motif analysis detected[GS],[DSD],and[S..E]as major phosphorylation sites in casein kinase-II and mitogen-activated protein kinases regulated by drought priming and heat stress.Functional annotation and gene ontology analysis demonstrated that DRPs in response to drought priming and in droughtprimed plants subsequently exposed to heat stress were mostly enriched in four major biological processes,including RNA splicing,transcription control,stress protection/defense,and stress perception/signaling.These results suggest the involvement of post-translational regulation of the aforementioned biological processes and signaling pathways in drought priming memory and cross-tolerance with heat stress in a temperate grass species.

关键词： phosphorylation drought temperate

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Associations of PGK1 promoter hypomethylation and PGK1-mediated PDHK1 phosphorylation with cancer stage and prognosis:a TCGA pan-cancer analysis

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Cancer Communications 2019年第1期39卷 482-498页

作者： Fei Shao Xueying Yang Wei Wang Juhong Wang Wei Guo Xiaoli Feng Susheng Shi Qi Xue Shugeng Gao Yibo Gao Zhimin Lu Jie He Department of Thoracic Surgery National Cancer Center/National Clinical Research Center for Cancer/Cancer HospitalChinese Academy of Medical Sciences and Peking Union Medical CollegeNo.17 PanjiayuannanliChaoyang DistrictBeijing 100021P.R.China Qingdao Cancer Institute Cancer Institute of The Affiliated Hospital of Qingdao UniversityQingdao 266003ShandongP.R.China Zhejiang Provincial Key Laboratory of Pancreatic Disease The First Affiliated Hospitaland Institute of Translational MedicineZhejiang University School of MedicineHangzhou 310029ZhejiangP.R.China Department of Pathology National Cancer Center/National Clinical Research Center for Cancer/Cancer HospitalChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100021P.R.China State Key Laboratory of Molecular Oncology National Cancer Center/National Clinical Research Center for Cancer/Cancer HospitalChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100021P.R.China

Background:Cancer cells reprogram metabolism for proliferation.Phosphoglycerate kinase 1(PGK1),as a glycolytic enzyme and newly identified protein kinase,coordinates glycolysis and mitochondrial metabolism.However,the clini-cal significance of PGK1 expression and function in cancer progression is unclear.Here,we investigated the relation-ship between the progression and prognosis of multiple cancer types and PGK1 expression and its function in the mitochondrial metabolism regulation.Methods:We performed pan-cancer analyses of PGK1 mRNA level and DNA methylation in 11,908 tumor tissues and 1582 paired normal tissues across 34 cancer types in The Cancer Genome Atlas datasets.Using specific antibodies against PGK1 S203 and PDHK1 T338 phosphorylation,we performed immunohistochemistry with tissue microarray assay in additional 818 cancer cases with 619 paired normal tissues from five cancer types.Results:The PGK1 mRNA level was significantly elevated with hypomethylation in promotor regions and associated with advanced TNM stage in 15 and four cancer types,respectively.In breast carcinoma,elevated PGK1 mRNA level and promoter hypomethylation were associated with poor prognosis.Positively correlated PGK1 S203 and PDHK1 T338 phosphorylation levels were significantly associated with short overall survival(OS)in cancers of the breast,liver,lung,stomach,and esophagus and with advanced TNM stage in breast and esophageal cancers.PGK1 pS203 and PDHK1 pT338 were also independent predictors of short OS in liver,lung,and stomach cancer.Conclusions:The elevated expression,promoter hypomethylation,and phosphorylation of PGK1 and PDHK1 were related with disease progression and short OS in diverse types of cancer.PGK1 and PDHK1 phosphorylation may be potential prognostic biomarkers.

关键词： PGK1 Cancer metabolism Epigenetics phosphorylation Methylation The Cancer Genome Atlas Pan-cancer analysis Prognosis Overall survival

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Simvastatin Increases the Activity of Endothelial Nitric Oxide Synthase via Enhancing phosphorylation

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2009年第3期29卷 286-290页

作者：李小霞汪培华徐西振王勇夏永汪道文 The Institute of Hypertension and Department of Internal Medicine Tongji Hospital Tongji Medical College Huazhong University of Science Davis Heart & Lung Research Institute the Ohio State University Medical Center

3-hydroxy-3-methylgulutaryl-coenzyme A （HMG-CoA） reductase inhibitors or statins are a kind of lipid-lowering agents and have been used for the prevention and treatment of Cardiovascular diseases. Recent studies suggested that statins, besides lowering cholesterol, may protect vessels by enhancing the activity of endothelial nitric oxide synthase （eNOS）. In the present study, we investigated if simvastatin increases eNOS activity through its phosphorylation in 293 cells （293-eNOS） with stable expression of eNOS. The results showed that incubation of 293-eNOS cells with simvastatin （10 μm/L） for 2 h significantly increased in the activity of eNOS as shown by the conversion of L-arginine to L-citrulline （2889.70±201.51 versus 5630.18＋218.75 pmol/min . mg proteins）（P〈0.01）. Western blotting revealed that simvastatin increased phosphorylation of eNOS at 1177 （ser） and also 495 （thr） but did not affect the overall expression of eNOS or inducible NOS. Further study found that simvastatin raised phosphorylation levels of Akt and AMPK, and such effect could be antagonized by Akt inhibitor or AMPK inhibitor. These results suggest that simvastatin could stimulate,the activity of eNOS via its phosphorylation by Akt and AMPK, which provides a new mechanism, other than lipid-lowering effect, for the cardiovascular protection of statins.

关键词： simvastatin nitric oxide synthase phosphorylation endothelial cells

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Changes in phosphorylation of Connexin43 in Rats during Acute Myocardial Hypoxia and Effects of Antiarrhythmic Peptide on the phosphorylation

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2007年第3期27卷 241-244页

作者：王荣张存泰阮燕菲刘念王琳 Department of Cardiology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology

In order to confirm the hypothesis that during acute hypoxia, the antiarrhythmic peptide (AAP10) could improve conductance by changing the phosphorylation state of connexin43 (Cx43), isolated perfused rat hearts were randomly divided into three groups: control, hypoxia and AAP10 (n=9 in each group). The change in Cx43 phosphorylation was tested by Western-blot; the distribu- tion of Cx43 was observed by confocal immunofluorescence microscopy. Western-blot analysis re- vealed that the expression of total Cx43 protein was significantly decreased during acute hypoxia, while nonphosphorylated Cx43 (NP-Cx43) was unchanged. AAP10 could increase the expression of total Cx43 protein, but had no effects on the NP-Cx43 protein. Immunofluorescence study showed that during acute hypoxia, both total Cx43 and NP-Cx43 proteins were greatly decreased, while AAP10 only increased the expression of total Cx43 protein, but had no effect of the NP-Cx43 protein expression. These findings suggested that the decrease of intercellular communication may be associ- ated with the reduction of phosphorylated Cx43 (p-Cx43) and translocation of NP-Cx43 from the surface of gap junction into intracellular pools during acute hypoxia. AAP10 can improve intercelluar communication by enhancing phosphorylation of Cx43.

关键词： hypoxia connexin43 phosphorylation antiarrhythmic peptide

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Reversal of Multidrug Resistance and Inhibition of phosphorylation of AKT in Human Ovarian Cancer Cell Line by Wild-type PTEN Gene

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2007年第6期27卷 713-716页

作者：吴卉娟翁丹卉邢辉卢运萍马丁 Department of Obstetrics and Gynecology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology

The reversing effect of wild-type PTEN gene on resistance of C 13K cells to cisplatin and its inhibitory effect on the phosphorylation of protein kinase B （AKT） were studied. The expression of PTEN mRNA and protein in OV2008 cells and C13K cells were semi-quantitatively detected by using RT-PCR and Western blotting. Recombinant eukaryotic expression plasmid containing human wild-type PTEN gene was transfected into C13K cells by lipofectamine2000. The expression of PTEN mRNA was monitored by RT-PCR and the expression of PTEN, Akt, p-Akt protein were ana- lyzed by Western blotting in PTEN-transfected and non-transfected C13K cells. Proliferation and chemosensitivity of cells to DDP were measured by MTT, and cell apoptosis was detected by flow cytometry after treatment with cisplatin. The expression of PTEN mRNA and protein in OV2008 cells were significantly higher than those in C13K cells. After transfection with PTEN gene for 48 h, the expression of PTEN mRNA and protein in C 13K cells were 2.04 ± 0.10, 0.94± 0.04 respectively and the expression of p-Akt protein （ 0.94± 0.07） was lower than those in control groups （1.68 ±0.14, 1.66± 0.10）（P〈 0.05）. The IC50 of DDP to C 13 K cells transfected with PTEN （7.2± 0.3 la mol/L） was obviously lower than those of empty-vector transfected cells and non-transfected cells （12.7±0.4 lamol/1, 13.0±0.3 lamol/L）（P〈0.05）. The apopototis ratio of wild-type PTEN-transfected, empty vector transfected and non-transfected C13K cells were （41.65___0.87）%, （18.61 ±0.70）% and （15.28±0.80）% respectively, and the difference was statistically significant （P〈0.05）. PTEN gene plays an important role in ovarian cancer multidrug resistance. Transfection of PTEN could increase the expression of PTEN and restore drug sensitivity to cisplatin in human ovarian cancer cell line C 13K with multidrug-resistance by decreasing the expression of p-Akt.

关键词： multidrug resistance phosphorylation AKT ovarian cancer cells wild-type PTEN gene

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Relationship between Tyrosine phosphorylation and Protein Expression of Insulin Receptor and Insulin Resistance in Gestational Diabetes Mellitus

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2014年第3期34卷 393-397页

作者：初永丽宫玉典苏志慧于洪娜崔青姜海洋曲洪美 Department of Obstetrics and Gynecology the Affiliated Yantai Yuhuangding Hospital of Qingdao University Medical College Yantai Psychological Rehabilitation Hospital

Summary： The relationship between tyrosine phosphorylation （TP） and protein expression of insulin receptor （InsR） and insulin resistance （IR） in patients with gestational diabetes mellitus （GDM） was investigated. The InsR expression and TP in skeleton muscle tissue were determined by Western blotting and immunoprecipitation in women with GDM （GDM group, n=22）, normal pregnant women （normal pregnancy group, n=22） and normal non-pregnant women （normal non-pregnant group, n=13）. Fasting plasma glucose （FPG） and fasting insulin （FINS） were measured by oxidase assay and immunoradioassay. The results showed that the levels of FPG （5.61±0.78 mmol/L）, FINS （15.42±5.13 mU/L） and Ho- meostasis model assessment-IR （HOMA-IR）（1.21±0.52） in GDM group were significantly higher than those in normal pregnancy group （4.43±0.46 mmol/L, 10.56±3.07 mU/L and 0.80±0.31 respectively）（P〈0.01）. The levels of FINS and HOMA-IR in normal pregnancy group were significantly higher than those in normal non-pregnant group （7.56±2.31 mU/L and 0.47±0.26 respectively）（P〈0.01）. There was no significant difference in the InsR expression level among the three groups （P〉0.05）. TP of InsR with insulin stimulation was significantly decreased in GDM group （0.20±0.05） as compared with normal pregnancy group （0.26±0.06）（P〈0.01）. TP of InsR with insulin stimulation in normal pregnancy group was lower than that in normal non-pregnant group （0.31±0.06）（P〈0.01）. TP of InsR with insulin stimu- lation was negatively related with HOMA-IR in GDM group （r=-0.525, P〈0.01）. There was no correlation between the protein expression of InsR and HOMA-IR in GDM group （r=-0.236, P〉0.05）. It was suggested that there is no significant correlation between the protein expression of InsR in skeletal muscle and IR in GDM, but changes in TP of InsR are associated with IR in GDM.

关键词： gestational diabetes mellitus insulin resistance tyrosine phosphoproteins phosphorylation

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Carboxyl terminal of rhodopsin kinase is required for the phosphorylation of photo-activated rhodopsin

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Cell Research 1998年第4期8卷 303-310页

作者： YU QING MING ZHI JIE CHENG JIAN ZHAO TIAN HUA ZHOU YA LAN WU LAN MA GANG PEI(Shanghai Institute of Cell Biology, Chinese Academy of Science, 320 Yue Yang Road, Shanghai 200031, China)(Department of Neurobiology, Shanghai Medical University,Shanghai Shanghai Institute of Cell Biology Chinese Academy of Science Shanghai China Department of Neurobiology Shanghai Medical University Shanghai China

Human rhodopsin kinase (RK) and a carboxyl terminus-truncated mutant RK lacking the last 59 amino acids (RKC) were expressed in human embryonic kidney 293 cells to investigate the role of the carboxyl terminus of RK in recognition and phosphorylation of rhodopsin. RKC, like the wild-type RK, was detected in both plasma membranes and cytosolic fractions. The Cterminal truncated rhodopsin kinase was unable to phosphorylate photo-activated rhodopsin, but possesses kinase activity similar to the wild-type RK in phosphorylation of small peptide substrate. It suggests that the truncation did not disturb the gross structures of RK catalytic domain. Our results also show that RKC failed to translocate to photo-activated rod out segments. Taken together,our study demonstrate the carboxyl terminus of RK is required for phosphorylation of photo-activated rhodopsin and strongly indicate that carboxyl-terminus of RK may be involved in interaction with photo-activated rhodopsin.

关键词： Rhodopsin kinase phosphorylation fruncation

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Protein phosphorylation and Phosphoproteome:An Overview of Rice

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Rice science 2020年第3期27卷 184-200页

作者： Abolore Adijat AJADI Amara CISSE Shakeel AHMAD WANG Yifeng SHU Yazhou LI Shufan LIU Xixi Babatunde Kazeem BELLO Sani Muhammad TAJO TONG Xiaohong ZHANG Jian State Key Laboratory of Rice Biology/China National Center for Rice Improvement/Laboratory of Rice Stress Physiology and Regulation China National Rice Research InstituteHangzhou 310006China Biotechnology Unit National Cereals Research InstituteBadeggiNiger State 912101Nigeria Lianyungang Academy of Agricultural Sciences Lianyungang 222243China Bioresources Development Center National Biotechnology Development AgencyKano 700241Nigeria

Protein phosphorylation,one of the major post-translational modifications,plays a crucial role in cell signaling,DNA replication,gene expression and differentiation;and alters enzyme activity and other biological activities;and regulates cell proliferation and enlargement,phytohormone biosynthesis and signaling,plant disease resistance,and grain filling and quality during rice seed development.Research work on protein phosphorylation started in the 1950 s with the discovery of phosphorylase a and phosphorylase b which are phospho and dephospho forms of the same enzyme.Over the last decade,rice proteomics has accomplished tremendous progress in setting up techniques to proteome nearly all tissues,organs and organelles.The progress made in this field is evident in number of research works.However,research on rice protein phosphorylation is still at its infancy and there are still many unanswered questions.In this review,the general description of protein phosphorylation,including history,structure,frequency of occurrence and function,are discussed.This work also elucidates the different methods for identification,qualification and finally,the progress in rice phosphoproteome research and perspectives.

关键词： phosphoproteome protein phosphorylation post-translational modification rice

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Endoplasmic reticulum stress is involved in okadaic acid-induced tau phosphorylation and neurotoxic effects of okadaic acid

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Neural Regeneration Research 2010年第20期5卷 1534-1540页

作者： Lijie Feng Yuxian Shen Aimin Sun Yujun Shen Shengyun Fang Jun Li School of Pharmacy Anhui Medical University Hefei 230032 Anhui Province China Key Laboratory of Gene Resource Utilization for Genetic Diseases of Educational Ministry and Anhui Province Anhui Medical University Hefei 230032 Anhui Province China Institute of Medical and Pharmaceutical Biotechnology Anhui Medical University Hefei 230032 Anhui Province China

The role of the endoplasmic reticulum in Alzheimer＇s disease pathogenesis remains poody understood.The present study investigated the correlation of okadaic acid-induced tau hyperphosphorylation and neurotoxicity with endoplasmic reticulum stress.Following co-culture of various concentrations of okadaic acid （25,50,100,and 200 nmol/L） and human neuroblastoma SH-SY5Y cells,okadaic acid,a selective protein phosphatase （PP-1,PP-2A） inhibitor,reduced cell viability in a concentration-dependent manner,which was closely associated with okadaic acid-induced tau phosphorylation.In addition,expressions of the unfolded protein response activation marker glucose-regulated protein 78 （BiP/Grp78） and the CCAAT/enhancer binding protein homologous protein （CHOP）/Gadd153 increased following okadaic acid treatment.Furthermore,the unfolded protein response was activated in human embryonic kidney 293 （HEK 293） cells overexpressing tau following okadaic acid treatment,but untransfected HEK 293 cells did not exhibit activation of the unfolded protein response.These data indicate that endoplasmic reticulum stress is involved in mechanisms underlying tau phosphorylation and the cytotoxic effects of okadaic acid.

关键词： tau phosphorylation endoplasmic reticulum stress unfolded protein response

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ATP regulates the phosphorylation and degradation of myofibrillar proteins in ground ovine muscle

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Journal of Integrative Agriculture 2021年第1期20卷 311-318页

作者： REN Chi HOU Cheng-li ZHANG De-quan LI Xin XIAO Xiong BAI Yu-qiang Institute of Food Science and Technology Chinese Academy of Agricultural Sciences/Key Laboratory of Agro-Products ProcessingMinistry of Agriculture and Rural AffairsBeijing 100193P.R.China

phosphorylation post-translational modification plays an important role in postmortem muscle quality traits. Adenosine triphosphate(ATP) is an energy source and a key substrate of phosphorylation which provides the phosphatase groups to proteins in the presence of protein kinases. However, in postmortem muscle, the effects of ATP content on phosphorylation are poorly studied. The study investigated the effect of ATP on protein phosphorylation and degradation in postmortem ovine muscle. The ground muscle with/without additional ATP were treated/control groups and stored at 25 and 4℃, respectively. The ATP content led to different changes of p H value between the ATP-treated and control groups. The phosphorylation level of myofibrillar proteins was higher(Pphosphorylation. A slower degradation rate of μ-calpain, desmin and troponin T was observed in the ATP-treated group which showed that there was a negative relationship between ATP level and the degradation of proteins. These observations clearly highlighted the role of ATP on the development of meat quality by regulating the phosphorylation and degradation of myofibrillar proteins in postmortem ovine muscle.

关键词： ATP postmortem ovine muscle phosphorylation protein degradation

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