The Role of the Charged Residues of the GP2 Helical Regions in Ebola Entry

查看全文 作　　者：Haiqing [1]Jiang;Jizhen [1]Wang;Balaji [1]Manicassamy;Santhakumar [1]Manicassamy;Michael [2]Caffrey;Lijun [1]Rong

机构地区：[1]Department of Microbiology and Immunology, College of Medicine, University of Illinois at Chicag;[2]Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago College of Medicine,Chicago, IL 60612, USA

出　　处：《Virologica Sinica》索引2009年第24卷第2期,共15页

基　　金：supported by National Institutes of Health grants CA 092459 and AI48056. L. R. was a recipient of the Schweppe Foundation Career Development Award. 摘　　要：The glycoprotein(GP) of Ebola is the sole structural protein that forms the spikes on the viral envelope. The GP contains two subunits,GP1 and GP2,linked by a disulfide bond,which are responsible for receptor binding and membrane fusion,respectively. In this study,the full length of GP gene of Ebola Zaire species,2028 base pairs in length,was synthesized using 38 overlapping oligonucleotides by multiple rounds of polymerase chain reaction(PCR) . The synthesized GP gene was shown to be efficiently expressed in mammalian cells. Furthermore,an efficient HIV-based pseudotyping system was developed using the synthetic GP gene,providing a safe approach to dissecting the entry mechanism of Ebola viruses. Using this pseudotyping system and mutational analysis,the role of the charged residues in the GP2 helical regions was examined. It was found that substitutions of the most charged residues in the regions did not adversely affect GP expression,processing,or viral incorporation,however,most of the mutations greatly impaired the ability of GP to mediate efficient viral infection. These results demonstrate that these charged residues of GP2 play an important role in GP-mediated Ebola entry into its host cells. We propose that these charged residues are involved in forming the intermediate conformation(s) of GP in membrane fusion and Ebola entry. 关键词：埃博拉病毒残留物入境螺旋糖蛋白基因聚合酶链反应哺乳动物细胞结构蛋白

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The Role of the Charged Residues of the GP2 Helical Regions in Ebola Entry

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