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eukaryotic initiation factor 5A2 and human digestive system neoplasms

World Journal of Gastrointestinal Oncology 2019年第6期11卷 449-458页

作者： Qing-Bin Meng Jing-Jing Peng Zi-Wei Qu Xiao-Min Zhu Zhang Wen Wei-Ming Kang Department of Gastrointestinal Surgery the First Hospital of Wuhan CityWuhan 430022Hubei ProvinceChina Department of Gastroenterology General Hospital of the Yangtze River ShippingWuhan 430015Hubei ProvinceChina 不详 Department of Hepato-Biliary-Pancreatic Surgery and Liver Transplantation the First Affiliated Hospital of Guangxi Medical UniversityNanning 530021Guangxi Zhuang Autonomous RegionChina Department of General Surgery Peking Union Medical College HospitalChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijing 100730China

eukaryotic initiation factor 5A2(eIF5A2),as one of the two isoforms in the family,is reported to be a novel oncogenic protein that is involved in multiple aspects of many types of human cancer.Overexpression or gene amplification of EIF5A2 has been demonstrated in many cancers.Accumulated evidence shows that eIF5A2 initiates tumor formation,enhances cancer cell growth,increases cancer cell metastasis,and promotes treatment resistance through multiple means,including inducing epithelial–mesenchymal transition,cytoskeletal rearrangement,angiogenesis,and metabolic reprogramming.Expression of eIF5A2 in cancer correlates with poor survival,advanced disease stage,as well as metastasis,suggesting that eIF5A2 function is crucial for tumor development and maintenance but not for normal tissue homeostasis.All these studies suggest that eIF5A2 is a useful biomarker in the prediction of cancer prognosis and serves as an anticancer molecular target.This review focuses on the expression,subcellular localization,post-translational modifications,and regulatory networks of eIF5A2,as well as its biochemical functions and evolving clinical applications in cancer,especially in human digestive system neoplasms.

关键词： eukaryotic translation initiation factor 5A2 Hypusine modification Acetylation modification Drug resistance Cancer therapeutics

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eukaryotic Translation Initiation Factors Shape RNA Viruses Resistance in Plants

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Horticultural Plant Journal 2020年第2期6卷 81-88页

作者： Jannat Shopan Xiaolong Lv Zhongyuan Hu Mingfang Zhang Jinghua Yang Laboratory of Germplasm Innovation and Molecular Breeding Institute of Vegetable ScienceZhejiang UniversityHangzhou 310058China Key Laboratory of Horticultural Plant Growth and Development Ministry of Agriculture and Rural AffairsHangzhou 310058China

Viruses are representative of a global threat to agricultural production. Genetic resistance is the preferred strategy for the control of viral infection and against loss of crop yield. Viral protein synthesis requires host cellular factors for translating their viral RNAs, and for regulating their replication and cell to cell systemic movement. Therefore, the viruses are dependent on cellular translation factors. Mutations in the gene encoding eIF4E and eIF4G or their isoforms, eIFiso4 E, eIFiso4 G and eIF2Bβ have been mapped as a source of plant potyvirus while other genus of plant virus recessive resistance genes in many species are originated from these loci. Some of other plant translation factors, such as eIF3,eIF4 A-like helicases, eEF1A and eEF1B, which are required in interacting with viral RNAs and regulating various aspects of the infection cycle,have also been identified. Here, we summarized the mechanisms utilized by RNA viruses of eukaryotic plants and the essential roles of e IFs in virus infection. Moreover, we discussed the potential of e IFs as a target gene in the development of genetic resistance to viruses for crop improvement. This review highlighted newly revealed examples of abnormal translational strategies and provided insights into natural host resistance mechanisms that have been linked to 3 cap-independent translational enhancer activity.

关键词： eukaryotic translation initiation factor genome editing 3 cap-independent translational enhancer virus resistance

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Construction and biological activities of human tPA eukaryotic expression plasmid

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Hepatobiliary & Pancreatic Diseases International 2004年第4期3卷 511-515页

作者： ZhongJun Wu SuFen Yang ShuSen Zheng De Shi WeiWei Wu DeWei Li Organ Transplant Center First Affiliated Hospital Zhejiang University School of Medicine Hangzhou 310003 China. wzjxdfpxy@***

BACKGROUND:Tissue-type plasminogen activator(tPA), which is highly efficient and specific in resolution of thrombus, could significantly improve the survival rate and life-quality of the patients with thrombosis. This study was designed to clone human tPA gene, construct eukaryotic expression plasmid pcDNA3.1(+)/tPA, and evaluate their biological activities.;METHODS:The tPA gene was obtained from dead human heart tissue with reverse transcriptase-polymerase chain reaction (RT-PCR). It was cloned to eukaryotic expression plasmid pcDNA3.1(+) by recombination strategy. The eukaryotic expression plasmid pcDNA3.1(+)/tPA was identified by restriction enzyme digestion and sequenced. The pcDNA3.1(+)/tPA was transfected into vascular smooth muscle cell (VSMC) by using lipofection. The tPA expression level was detected by Northern blot and dot blot, and the protein biological activities of tPA were detected by the fibrin plate technique.;RESULTS:The tPA gene was cloned and pcDNA3.1(+)/tPA was constructed. The tPA expression levels of mRNA and protein were highly increased after pcDNA3.1(+)/tPA transfected into VSMC, and the expression of tPA protein showed evident biological activities.;CONCLUSIONS:The present study has laid a foundation for further animal experiment in treating thrombus in transplanted organ by tPA gene transfection.

关键词： tissue-type plasminogen activator eukaryotic expression plasmid vascular endothelial cell organ transplantation

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Construction of eukaryotic expression plasmids of hepatitis B surface antigen and helper T lymphocyte epitope

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Hepatobiliary & Pancreatic Diseases International 2004年第2期3卷 219-222页

作者： YunFeng Piao TongYu Tang JunQi Niu Feng Wang Department of Gastroenterology First Hospital of Jilin University Changchun 130021 China.

BACKGROUND:DNA immunization provides a promising approach to elicit protective humoral and cellular immune responses against HBV. This study was to construct an eukaryotic expression plasmid containing helper T lymphocyte epitope, which will enhance the immunogenicity of a novel hepatitis B virus (HBV) fusion protein DNA vaccine.;METHODS:The target gene containing pan-DR helper T cell epitopes (PADRE) and HBsAg was amplified by polymerase chain reaction (PCR). The PCR products were linked with PMD-18T vector. Plasmid DNA was purified from transformed E.coli competent cell JM109 and digested with Hind III and EcoR I. Then, the target gene was cloned in pcDNA3.1(+) digested by Hind III and EcoR I. Finally, the identity of DNA was verified by digestion and DNA sequencing.;RESULTS:The recombinant expression vectors of pcDNA3.1(+)-PADRE/HBs were identified by restriction enzyme digestion and DNA sequencing. The insert DNA fragment was consistent with the expected sequence.;CONCLUSIONS:The constructed eukaryotic expression plasmid of pcDNA3.1(+)-PADRE/HBs is convenient for further study of eukaryotic transfection and response for cellular and humoral immunity against HBV.

关键词： hepatitis B virus helper T lymphocyte epitope eukaryotic expression plasmid

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eukaryotic elongation factor-1α 2 knockdown inhibits hepatocarcinogenesis by suppressing PI3K/Akt/NF-κB signaling

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World Journal of Gastroenterology 2016年第16期22卷 4226-4237页

作者： Fu-Nan Qiu Yi Huang Dun-Yan Chen Feng Li Yan-An Wu Wen-Bing Wu Xiao-Li Huang Provincial Clinical College Fujian Medical University Department of Hepatobiliary Surgery Fujian Provincial Hospital Department of Clinical Laboratory Fujian Provincial Hospital Department of Pathology Fujian Provincial Hospital

AIM: To assess the impact of eukaryotic elongation factor 1 alpha 2(e EF1A2) on hepatocellular carcinoma(HCC) cell proliferation, apoptosis, migration and invasion, and determine the underlying mechanisms.METHODS: e EF1A2 levels were detected in 62 HCC tissue samples and paired pericarcinomatous specimens, and the human HCC cell lines SK-HEP-1, Hep G2 and BEF-7402, by real-time PCR and immunohistochemistry. Experimental groups included e EF1A2 silencing in BEL-7402 cells with lentivirus e EF1A2-sh RNA(KD group) and e EF1A2 overexpression in SK-HEP-1 cells with e EF1A2 plasmid(OE group). Non-transfected cells(control group) and lentivirusbased empty vector transfected cells(NC group) were considered control groups. Cell proliferation(MTT and colony formation assays), apoptosis(Annexin V-APC assay), cell cycle(DNA ploidy assay), and migration and invasion(Transwell assays) were assessed. Protein levels of PI3K/Akt/NF-κB signaling effectors were evaluated by Western blot.RESULTS: e EF1A2 m RNA and protein levels were significantly higher in HCC cancer tissue samples than in paired pericarcinomatous and normal specimens. SK-HEP-1 cells showed lower e EF1A2 m RNA levels; Hep G2 and BEL-7402 cells showed higher e EF1A2 m RNA levels, with BEL-7402 cells displaying the highest amount. Efficient e EF1A2 silencing resulted in reduced cell proliferation, migration and invasion, increased apoptosis, and induced cell cycle arrest. The PI3K/Akt/NF-κB signaling pathway was notably inhibited. Inversely, e EF1A2 overexpression resulted in promoted cell proliferation, migration and invasion.CONCLUSION: e EF1A2, highly expressed in HCC, is a potential oncogene. Its silencing significantly decreases HCC tumorigenesis, likely by inhibiting PI3K/Akt/NF-κB signaling.

关键词： Hepatocellular carcinoma Carcinogenesis eukaryotic elongation factor 1 alpha 2 Proliferation PI3K/Ak

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Cellular Caspase-3 Contributes to EV-A71 2Apro-Mediated Down-Regulation of IFNAR1 at the Translation Level

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Virologica Sinica 2020年第1期35卷 64-72页

作者： Bangtao Chen Yuya Wang Xinyi Pei Sanyuan Wang Hao Zhang Yihong Peng Department of Microbiology&Infectious Disease Center School of Basic Medical SciencesPeking University Health Science CenterBeijing 100083China

Enterovirus A71(EV-A71) is the major pathogen responsible for the severe hand, foot and mouth disease worldwide, for which few effective antiviral drugs are presently available. Interferon-a(IFN-a) has been used in antiviral therapy for decades;it has been reported that EV-A71 antagonizes the antiviral activity of IFN-a based on viral 2 Apro-mediated reduction of the interferon-alpha receptor 1(IFNAR1);however, the mechanism remains unknown. Here, we showed a significant increase in IFNAR1 protein induced by IFN-a in RD cells, whereas EV-A71 infection caused obvious downregulation of the IFNAR1 protein and blockage of IFN-a signaling. Subsequently, we observed that EV-A71 2 Apro inhibited IFNAR1 translation by cleavage of the eukaryotic initiation factor 4 GI(eIF4GI), without affecting IFNAR1 m RNA levels induced by IFN-a. The inhibition of IFNAR1 translation also occurred in puromycin-induced apoptotic cells when caspase-3 cleaved e IF4 GI. Importantly, we verified that 2 Aprocould activate cellular caspase-3, which was subsequently involved in e IF4 GI cleavage mediated by 2 Apro. Furthermore, inhibition of caspase-3 activation resulted in the partial restoration of IFNAR1 in cells transfected with 2 A or infected with EV-A71, suggesting the pivotal role of both viral 2 Aproand caspase-3 activation in the disturbance of IFN-a signaling. Collectively, we elucidate a novel mechanism by which cellular caspase-3 contributes to viral 2 Apro-mediated down-regulation of IFNAR1 at the translation level during EV-A71 infection, indicating that caspase-3 inhibition could be a potential complementary strategy to improve clinical anti-EV-A71 therapy with IFN-a.

关键词： Enterovirus A71(EV-A71) Interferon alpha receptor 1(IFNAR1) 2A protease(2Apro) Caspase-3 eukaryotic initiation factor 4GI(eIF4GI)

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Effect of siRNA-mediated knockdown of eIF3c gene on survival of colon cancer cells

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2013年第6期14卷 451-459页

作者： Ning SONG Yan WANG Xiao-dong GU Zong-you CHEN Liu-bin SHI Department of General Surgery Huashan HospitalFudan University

eukaryotic initiation factor subunit c(eIF3c) has been identified as an oncogene that is over-expressed in tumor cells and,therefore,is a potential therapeutic target for gene-based cancer treatment.This study was focused on investigating the effect of small interfering RNA(siRNA)-mediated eIF3c gene knockdown on colon cancer cell survival.The eIF3c gene was observed to be highly expressed in colon cancer cell models.The expression levels of the gene in eIF3c siRNA infected and control siRNA infected cells were compared via real-time polymerase chain reaction(PCR) and western blotting analysis.Cell proliferation levels were analyzed employing 3-(4,5-dimethylthiazol 2-yl)-2,5-diphenyltetrazolium bromide(MTT) and colony formation assays.Furthermore,the effects of eIF3c gene knockdown on the cell cycle and apoptosis were analyzed using flow cytometry.The results showed that suppression of eIF3c expression significantly(P<0.001) reduced cell proliferation and colony formation of RKO colon cancer cells.The cell cycle was arrested by decreasing the number of cells entering S phase.Further,apoptosis was induced as a result of eIF3c knockdown.Collectively,eIF3c deletion effectively reduced the survival of colon cancer cells and could be used as a therapeutic tool for colon cancer therapy.

关键词： eukaryotic initiation factor subunit c(eIF3c) Colon cancer RKO cells Small interfering RNA(siRNA)

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Localization and function of a eukaryotic-initiation-factor-2-associated 67-kDa glycoprotein

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World Journal of Biological Chemistry 2010年第10期1卷 313-320页

作者： Shiyong Wu,Edison Biotechnology Institute,Department of Chemistry and Biochemistry,Ohio University,Athens,OH 45701,United States Shiyong Wu Edison Biotechnology Institute Department of Chemistry and Biochemistry Ohio University Athens OH 45701 United States.

AIM: To study the localization and function of a eukaryotic initiation factor 2 (eIF2α)-associated 67-kDa glycoprotein (p67).METHODS: Immunofluorescence staining,35S-Met/Cys metabolic labeling,Western blotting analysis,sucrose gradient centrifugation and high speed centrifugation were used to determine the localization of proteins in transiently transfected COS-1 cells.Transient co-transfection followed by co-immunoprecipitation was used to study the interaction between p67 and double-stranded RNA (dsRNA)-dependent protein kinase (PKR).Wheat germ agglutinin agarose beads were used to absorb glycosylated proteins.In vivo 32P-labeling followed by immunoprecipitation and Western blotting were used to measure PKR autophosphorylation,eIF2α phosphorylation,and p67 expression in normal and breast cancer cells.RESULTS: The image from immunofluorescence staining showed that p67 was overexpressed in the cytosol but not in the nucleus.In a sucrose gradient,approxi-mately 30% of the overexpressed p67 was bound with ribosomes.p67 interacted with the kinase domain,butnot the dsRNA-binding domains of PKR.Only the glycosylated p67 was associated with the ribosome,and p67 did not compete with PKR for ribosome binding.In breast cancer cells,there was increased autophosphorylation of PKR but no phosphorylation of eIF2α,compared with normal breast cells.α The ratio of glycosylated/deglycosylated p67 was altered in breast cancer cells.CONCLUSION: Glycosylation of p67 is required for its ribosomal association and can potentially inhibit PKR via interaction with the kinase domain of PKR.

关键词： eukaryotic translation initiation factor 2 p67 Double-stranded RNA dependent protein kinase Phosphorylation Cancer

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Nuclear actin switch of the IN080 remodeler

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Journal of Molecular Cell Biology 2019年第5期11卷 343-344页

作者： Jun Wu Yimin Lao Bing Li Department of Biochemistry and Molecular Cell Biology Shanghai Jiao Tong University School of Medicine Shanghai 200025 China Department of Laboratory Medicine Shanghai General Hospital Shanghai Jiao Tong University Shanghai 200080 China

eukaryotic DNA is wrapped in nucleosomes, which impede the access of transcription factors and regulatory proteins to template DNA. Chromatin remodelers utilize the energy from ATP hydrolysis to drive histone movement relative to nucleosomal DNA and nucleosome editing. Thus, they play critical roles in transcription, DNA replication, and damage repair, and their dysfunctions are often associated with diseases including cancers (Klages-Mundt et al., 2018). Chromatin remodelers can be generally categorized into IN080, SWI/SNF, CHD, and ISWI subfamilies, which share conserved catalytic ATPasetranslocase motors. However, how other auxiliary components of these multi-subunit machinery control their genomic recruitment and actions of DNA translocation remains as a major challenge of the field.

关键词： eukaryotic DNA nucleosomes of the field

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Phosphatized biotas from the Neoproterozoic Doushantuo Formation on the Yangtze Platform

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Chinese Science Bulletin 2002年第22期47卷 1918-1924页

作者： ZHOU Chuanming YUAN Xunlai XIAO Shuhai Nanjing Institute of Geology and Paleontology Chinese Academy of SciencesNanjing 210008China Department of Geology TWane UniversityNew OrleansLA 70118USA

The Doushantuo Formation in South China was deposited after the Nantuo glaciation but before the evolution of complex Ediacaran metazoans. It contains multiple taphonomic windows, in its cherts, carbonaceous shales, and phosphorites, onto the late Neoproterozoic biosphere. The phosphatic window is unusually clear; Doushantuo phosphorites at Weng’an (Guizhou Province) are known to exquisitely preserve a multitude of single-celled eukaryotes, muiticellular algae, and microscopic animals. Our recent survey reveals that, in addition to the now famous Weng’an locality, Doushantuo phosphorites at Baokang (Hubei), Chadian (Shaanxi), and Shangrao (Jiangxi) also contain diverse eukaryotes preserved at the cellular level. All these phosphorites were deposited in shallow-water environments, typically above fair weather wave base and close to ancient islands. Along with Doushantuo cherts and shales, these phosphorites give us a clearer and more complete picture of late Neoproterozoic biological evolution: there

关键词： Neoproterozoic Doushantuo Formation phosphorite phosphatized biota eukaryotic fossils Yangtze Platform.

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