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O-GlcNAcylation regulates phagocytosis by promoting ezrin localization at the cell cortex

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Journal of Genetics and Genomics 2023年第7期50卷 486-496页

作者： Song Yang Hanyu Liu Hua Ni Lingyu Jiang Mulin Yang Quan Chen Jun Zhou Fan Yu State Key Laboratory of Medicinal Chemical Biology Frontiers Science Center for Cell ResponsesTianjin Key Laboratory of Protein ScienceHaihe Laboratory of Cell EcosystemCollege of Life SciencesNankai UniversityTianjin 300071China CNBG-Nankai Joint Research Center Nankai UniversityTianjin 300071China College of Life and Geographic Sciences Kashi UniversityXinjiang 844000China

O-GlcNAcylation is a post-translational modification that serves as a cellular nutrient sensor and participates in multiple physiological and pathological processes.However,it remains uncertain whether O-GlcNAcylation is involved in the regulation of phagocytosis.Here,we demonstrate a rapid increase in protein OGlcNAcylation in response to phagocytotic stimuli.Knockout of the O-GlcNAc transferase or pharmacological inhibition of O-GlcNAcylation dramatically blocks phagocytosis,resulting in the disruption of retinal structure and function.Mechanistic studies reveal that the O-GlcNAc transferase interacts with ezrin,a membrane-cytoskeleton linker protein,to catalyze its O-GlcNAcylation.Our data further show that ezrin OGlcNAcylation promotes its localization to the cell cortex,thereby stimulating the membrane-cytoskeleton interaction needed for efficient phagocytosis.These findings identify a previously unrecognized role for protein O-GlcNAcylation in phagocytosis with important implications in both health and diseases.

关键词： O-GlcNAcylation Phagocytosis Retinal pigment epithelium ezrin Cytoskeleton

Peptide targeting the interaction of S protein cysteine-rich domain with ezrin restricts pan-coronavirus infection

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Signal Transduction and Targeted Therapy 2023年第2期8卷 361-364页

作者： Zhuanchang Wu Xiaobo Lei Xin Wang Zhaoying Zhang Yuming Li Lifen Gao Xiaohong Liang Peihui Wang Jianwei Wang Chunhong Ma Key Laboratory for Experimental Teratology of Ministry of Education Key Laboratory of Infection and Immunity of Shandong Province and Dept.ImmunologySchool of Basic Medical SciencesCheeloo Medical CollegeShandong University250012JinanShandongChina NHC Key Laboratory of Systems Biology of Pathogens Institute of Pathogen BiologyChinese Academy of Medical Sciences and Peking Union Medical College100730BeijingChina College of Agriculture and Forestry Linyi UniversityLinyiShandongChina School of Public Health Shandong First Medical University&Shandong Academy of Medical Sciences271016Tai’anChina Key Laboratory for Experimental Teratology of Ministry of Education and Advanced Medical Research Institute Shandong University250012JinanShandongChina

Dear Editor,To date,seven human coronaviruses(HCoVs)have been identified,among which the highly pathogenic severe acute respiratory syndrome-associated coronavirus(SARS-CoV),Middle East respiratory syndrome coronavirus(MERS-CoV),and severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)have caused public health disasters worldwide.

关键词： respiratory acute ezrin

ezrin promotes invasion and migration of the MG63 osteosarcoma cell

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Chinese Medical Journal 2014年第10期127卷 1954-1959页

作者： Zhang Jian Zuo Jianhong Lei Mingsheng Wu Song Zang Xiaofang Zhang Chaoyue Department of Orthopaedics Third Xiangya Hospital Central SouthUniversity Changsha Hunan 421001 China Medical School University of South China Changsha Hunan421001 China People's Hospital of Zhangjiajie Zhangjiajie Hunan 427000 China

Background Evidence shows that ezrin plays an important role in the development of some human malignancies.But the mechanism by which ezrin may affect tumor cell invasion and metastasis remains unclear.Methods In this study,the expression of ezrin was verified in osteosarcoma （OS） cells and tissues by comparison with normal bone cells and tissues using Western blotting.OS-MG63 were transfected with pcDNA3.1-ezrin or pGenesil-1/ shRNA-ezrin and the stably transfected cells were selected with G418 to yield the ezrin cell line.The OS-MG63 tumor cells were delivered by tail vein to female BALB/c to develop pulmonary metastasis model in vivo.ezrin was identified as a direct target of miR-183 via a luciferase reporter carrying the 3＇-untranslated region of ezrin.Migration assays and invasion assays were done with the transwells.Signaling pathway was studied by Western blotting and/or inhibitor.Results Ectopic overexpression of ezrin in OS cell line MG63 promoted tumor cell invasion and migration.Consistent with this,knockdown of ezrin inhibited tumor cell invasion and migration.Similar results were obtained in the experimental metastasis model in vivo.We identified ezrin as a direct target of miR-183.What is more,ectopic expression of ezrin could induce the expression of N-cadherin and enhance the activity of extracellular signal-regulated kinase （ERK） signaling.Conclusion Collectively,these results suggest that ezrin as a direct target of miR-183 promotes the aggressiveness of OS via increased N-cadherin and activating ERK signaling.

关键词： osteosarcoma ezrin migration invasion N-cadherin

ezrin Promotes the Proliferation, Migration, and Invasion of Ovarian Cancer Cells

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Biomedical and Environmental Sciences 2021年第2期34卷 139-151页

作者： LI Mo Juan XIONG Dan HUANG Hao WEN Zhong Yong Department of Obstetrics and Gynecology Nanhai Hospital Affiliated to Southern Medical UniversityFoshan 528200GuangdongChina Department of Hematology Shunde Hospital of Southem Medical University(The First People's Hospital of Shunde)Foshan 528308GuangdongChina

Objective The underlying mechanism of ezrin in ovarian cancer(OVCA) is far from being understood.Therefore, this study aimed to assess the role of ezrin in OVCA cells(SKOV3 and Ca OV3) and investigate the associated molecular mechanisms.Methods We performed Western blotting, reverse transcription-quantitative polymerase chain reaction, MTT, cell colony, cell wound healing, transwell migration and invasion, Rho A and Rac active pull down assays, and confocal immunofluorescence experiments to evaluate the functions and molecular mechanisms of ezrin overexpression or knockdown in the proliferation and metastasis of OVCA cells.Results The ectopic expression of ezrin significantly increased cell proliferation, invasiveness, and epithelial–mesenchymal transition(EMT) in OVCA cells. By contrast, the knockdown of endogenous ezrin prevented OVCA cell proliferation, invasiveness, and EMT. Lastly, we observed that ezrin can positively regulate the active forms of Rho A rather than Rac-1 in OVCA cells, thereby promoting robust stress fiber formation.Conclusion Our results indicated that ezrin regulates OVCA cell proliferation and invasiveness by modulating EMT and induces actin stress fiber formation by regulating Rho-GTPase activity, which provides novel insights into the treatment of the OVCA.

关键词： Epithelial ovarian cancer ezrin Proliferation Invasiveness RhoA Stress fiber

The role of ezrin-associated protein network in human sperm capacitation

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Asian Journal of Andrology 2010年第5期12卷 667-676页

作者： Lei Wang Wen Chen Chun Zhao Ran Huo Xue-Jiang Guo Min Lin Xiao-Yan Huang Yun-Dong Mao Zuo-Min Zhou Jia-Hao Sha Laboratory of Reproductive Medicine Department of Histology and Embryology Nanjing Medical University Nanjing 210029 China Department of Reproductive Medicine the First Affiliated Hospital of Nanjing Medical University Nanjing 210029 China

Membrane modifications in sperm cells represent a key step in sperm capacitation; however, the molecular basis of these modifications is not fully understood. ezrin is the best-studied member of the ezrin/radixin/merlin family. As a cross-linker between the cortical cytoskeleton and plasma membrane proteins, ezrin contributes to remodeling of the membrane surface structure. Furthermore, activated ezrin and the Rho dissociation inhibitor, RhoGDI, promote the formation of cortical cytoskeleton-polymerized actin through Rho activation. Thus, ezrin, actin, RhoGDI, Rho and plasma membrane proteins form a complicated network in vivo, which contributes to the assembly of the structure of the membrane surface. Previously, we showed that ezrin and RhoGDI1 are expressed in human testes. Thus, we sought to determine whether the ezrin-RhoGDIl-actin-membrane protein network has a role in human sperm capacitation. Our results by Western blot indicate that ezrin is activated by phosphorylation of the threonine567 residue during capacitation. Co-immunoprecipitation studies revealed that, during sperm capacitation, the interaction between ezrin and RhoGDI1 increases, and phosphostaining of two dimensional electrophoresis gels showed that RhoGDI 1 is phosphorylated, suggesting that RhoGDI 1 dissociates from RhoA and leads to actin polymerization on the sperm head. We speculate that activated ezrin interacts with polymerized actin and the glycosylated membrane protein cd44 after capacitation. Blocking sperm capacitation using ezrin- or actin-specific monoclonal antibodies decreases their acrosome reaction （AR） rate, but has no effect on the AR alone. Taken together, our results show that a network consisting of ezrin, RhoGDI1, RhoA, F-actin and membrane proteins functions to influence the modifications that occur on the membrane of the sperm head during human sperm capacitation.

关键词： ezrin membrane sperm capacitation

ezrin调控肝癌细胞侵袭性伪足的形成促进肝细胞癌侵袭转移

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陆军军医大学学报 2022年第4期44卷 346-355页

作者：郑博文王保林路遥黄登张航刘嘉龙郑树国陆军军医大学(第三军医大学)第一附属医院全军肝胆外科研究所重庆400038 中国人民解放军西藏军区总医院肝胆科拉萨850032

目的探讨ezrin在肝癌细胞侵袭性伪足形成过程中的作用。方法利用GEPIA数据库分析肝癌组织中ezrin mRNA与伪足标志物Cortactin mRNA的表达相关性。将收集的105例肝细胞癌患者的组织标本制作成组织芯片,采用免疫组化检测ezrin蛋白和伪足... 详细信息

目的探讨ezrin在肝癌细胞侵袭性伪足形成过程中的作用。方法利用GEPIA数据库分析肝癌组织中ezrin mRNA与伪足标志物Cortactin mRNA的表达相关性。将收集的105例肝细胞癌患者的组织标本制作成组织芯片,采用免疫组化检测ezrin蛋白和伪足标志物Cortactin蛋白在肝癌组织中的表达,并分析二者的表达相关性。分析ezrin的表达水平与肝癌患者临床病理特征及预后的关系。敲低和上调肝癌细胞中ezrin的表达,采用Transwell实验观察ezrin蛋白的表达变化对肝癌细胞侵袭迁移能力的影响,采用Western blot和细胞免疫荧光技术检测ezrin蛋白表达变化对肝癌细胞侵袭性伪足形成能力的影响。结果生物信息学分析结果提示肝癌组织中ezrin mRNA与伪足标志物Cortactin mRNA的表达呈正相关(r=0.420,Pezrin蛋白与伪足标志物Cortactin蛋白的表达水平呈正相关(r=0.491,Pezrin蛋白的高表达与肝癌患者的血管侵犯率(P=0.016)及肿瘤相关死亡率(P=0.018)相关,且ezrin蛋白高表达患者的术后5年生存率及术后5年无瘤生存率均较低(Pezrin蛋白的表达可促进肝癌细胞的侵袭和迁移(Pezrin蛋白表达可抑制肝癌细胞侵袭性伪足的形成,并降低肝癌细胞的侵袭和迁移能力。结论ezrin可通过调控肝癌细胞侵袭性伪足的形成增强肝癌细胞的侵袭、迁移能力,进而促进肝癌进展。

关键词： ezrin Cortactin 肝细胞癌侵袭性伪足侵袭转移

ezrin： a regulator of actin microfilaments in ce unctions of the rat testis

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Asian Journal of Andrology 2015年第4期17卷 653-658页

作者： N Ece Gungor-Orduer Ciler Celik-Ozenci C Yan Cheng The Mary M. Wohlford Laboratory for Male Contraceptive Research Center for Biomedical Research Population Council New York USA Department of Histology andEmbryology Faculty of Medicine Akdeniz University Antalya Turkey.

ezrin, radixin, moesin and merlin （ERM） proteins are highly homologous actin-binding proteins that share extensive sequence similarity with each other. These proteins tether integral membrane proteins and their cytoplasmic peripheral proteins （e.g., adaptors, nonreceptor protein kinases and phosphatases） to the microfilaments of actin-based cytoskeleton. Thus, these proteins are crucial to confer integrity of the apical membrane domain and its associated junctional complex, namely the tight junction and the adherens junction. Since ectoplasmic specialization （ES） is an F-actin-rich testis-specific anchoring junction-a highly dynamic ultrastructure in the seminiferous epithelium due to continuous transport of germ cells, in particular spermatids, across the epithelium during the epithelial cycle-it is conceivable that ERM proteins are playing an active role in these events. Although these proteins were first reported almost 25 years and have since been extensively studied in multiple epithelia/endothelia, few reports are found in the literature to examine their role in the actin filament bundles at the ES. Studies have shown that ezrin is also a constituent protein of the actin-based tunneling nanotubes （TNT） also known as intercellular bridges, which are transient cytoplasmic tubular ultrastructures that transport signals, molecules and even organelles between adjacent and distant cells in an epithelium to coordinate cell events that occur across an epithelium. Herein, we critically evaluate recent data on ERM in light of recent findings in the field in particular ezrin regarding its role in actin dynamics at the ES in the testis, illustrating additional studies are warranted to examine its physiological significance in spermatogenesis.

关键词： blood-testis barrier ectoplasmic specialization ezrin spermatogenesis testis

Expression of ezrin,HGF,C-met in pancreatic cancer and non-cancerous pancreatic tissues of rats

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Hepatobiliary & Pancreatic Diseases International 2010年第6期9卷 639-644页

作者： Tan, Xing-Guo Yang, Zhu-Lin Cent S Univ Xiangya Hosp 2 Res Lab Hepatobiliary Dis Changsha 410011 Hunan Peoples R China

BACKGROUND: Recent studies have confirmed that the expression of ezrin, hepatocyte growth factor (HGF) and its receptor (C-met) is related to the genesis, progress, invasion and metastasis of some malignant tumors. Researches have also found that the biological function of ezrin is closely related to HGF/C-met in malignant tumors. However, there is no report on the expression levels of ezrin, HGF and C-met in rat pancreatic cancer induced by dimethylbenzanthracene (DMBA). This study aimed to detect the expression of ezrin, HGF and C-met in rat pancreatic cancer and non-cancerous pancreatic tissues, and assess its effect in cancer induction by DMBA. METHODS: Ninety Sprague-Dawley rats were divided into 3 groups randomly: 40 in a pancreatic cancer model group (group A), 40 in a trichostatin A (TSA) intervention group (group B), and 10 in a control group (group C). DMBA was directly implanted into the parenchyma of rat pancreas in group A+group B. The rats of group B were treated with 1 ml of TSA saline solution (1 mu g/ml) via intraperitoneal injection weekly. The carcinogenesis of rats executed within 3-5 months in groups A and B was observed by macrograph and microscopy. Meanwhile, the rats in group C were executed within 5 months. The EnVision (TM) immunohistochemistry for detecting the expression levels of ezrin, HGF and C-met was used in paraffin-embedded sections of the pancreatic specimens. RESULTS: The incidence of pancreatic cancer in group A was 48.6% and in group B 33.3%. The maximal diameter of tumor mass was significantly larger in group A than that in group B (Pezrin, HGF and C-met were significantly higher in ductal adenocarcinoma than in non-cancerous pancreatic tissues of groups A and B (Pezrin, HGF and C-met were significantly higher in ductal adenocarcinoma of group A than those in non-cancerous pancreatic tissues of group A (P<0.05)

关键词： pancreatic neoplasms animal model ezrin hepatocyte growth factor C-met

Acetylation of ezrin regulates membrane–cytoskeleton interaction underlying CCL18-elicited cell migration

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Journal of Molecular Cell Biology 2020年第6期12卷 424-437页

作者： Xiaoyu Song Wanjuan Wang Haowei Wang Xiao Yuan Fengrui Yang Lingli Zhao McKay Mullen Shihao Du Najdat Zohbi Saravanakumar Muthusamy Yalei Cao Jiying Jiang Peng Xia Ping He Mingrui Ding Nerimah Emmett Mingming Ma Quan Wu Hadiyah-Nicole Green Xia Ding Dongmei Wang Fengsong Wang Xing Liu School of Traditional Medicine Beijing University of Chinese MedicineBeijingChina MOE Key Laboratory for Membraneless Organelles&Cellular Dynamics Hefei National Center for Physical Sciences at the MicroscaleHefeiChina Morehouse School of Medicine Keck Center for Organoids PlasticityAtlantaGAUSA Optics and Optical Engineering University of Science and Technology of ChinaHefeiChina School of Life Science Anhui Medical UniversityHefeiChina

ezrin,a membrane–cytoskeleton linker protein,plays an essential role in cell polarity establishment,cell migration,and division.Recent studies show that ezrin phosphorylation regulates breast cancer metastasis by promoting cancer cell survivor and promotes intrahepatic metastasis via cell migration.However,it was less characterized whether there are additional post-translational modifications and/or post-translational crosstalks on ezrin underlying context-dependent breast cancer cell migration and invasion.Here we show that ezrin is acetylated by p300/CBP-associated factor(PCAF)in breast cancer cells in response to CCL18 stimulation.ezrin physically interacts with PCAF and is a cognate substrate of PCAF.The acetylation site of ezrin was mapped by mass spectrometric analyses,and dynamic acetylation of ezrin is essential for CCL18-induced breast cancer cell migration and invasion.Mechanistically,the acetylation reduced the lipid-binding activity of ezrin to ensure a robust and dynamic cycling between the plasma membrane and cytosol in response to CCL18 stimulation.Biochemical analyses show that ezrin acetylation prevents the phosphorylation of Thr567.Using atomic force microscopic measurements,our study revealed that acetylation of ezrin induced its unfolding into a dominant structure,which prevents ezrin phosphorylation at Thr567.Thus,these results present a previously undefined mechanism by which CCL18-elicited crosstalks between the acetylation and phosphorylation on ezrin control breast cancer cell migration and invasion.This suggests that targeting PCAF signaling could be a potential therapeutic strategy for combating hyperactive ezrin-driven cancer progression.

关键词： ezrin acetylation phosphorylation actin cell migration