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Defining the N-Linked Glycosylation Site of Hantaan Virus Envelope Glycoproteins Essential for Cell Fusion  

Zheng, Feng (Qilu Hospital of Shandong University)
Ma, Lixian (Qilu Hospital of Shandong University)
Shao, Lihua (School of Public Health of Shandong University)
Wang, Gang (Qilu Hospital of Shandong University)
Chen, Fengzhe (Qilu Hospital of Shandong University)
Zhang, Ying (Qilu Hospital of Shandong University)
Yang, Song (Qilu Hospital of Shandong University)
Publication Information
Journal of Microbiology / v.45, no.1, 2007 , pp. 41-47 More about this Journal
Abstract
The Hantaan virus (HTNV) is an enveloped virus that is capable of inducing low pH-dependent cell fusion. We molecularly cloned the viral glycoprotein (GP) and nucleocapsid (NP) cDNA of HTNV and expressed them in Vero E6 cells under the control of a CMV promoter. The viral gene expression was assessed using an indirect immunofluorescence assay and immunoprecipitation. The transfected Vero E6 cells expressing GPs, but not those expressing NP, fused and formed a syncytium following exposure to a low pH. Monoclonal antibodies (MAbs) against envelope GPs inhibited cell fusion, whereas MAbs against NP did not. We also investigated the N-linked glycosylation of HTNV GPs and its role in cell fusion. The envelope GPs of HTNV are modified by N-linked glycosylation at five sites: four sites on G1 (N134, N235, N347, and N399) and one site on G2 (N928). Site-directed mutagenesis was used to construct eight GP gene mutants, including five single N-glycosylation site mutants and three double-site mutants, which were then expressed in Vero E6 cells. The oligosaccharide chain on residue N928 of G2 was found to be crucial for cell fusion after exposure to a low pH. These results suggest that G2 is likely to be the fusion protein of HTNV.
Keywords
Hantaan virus; cell-cell fusion; glycoproteins; N-linked glycosylation;
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