scholarly journals An External Loop Region of Domain III of Dengue Virus Type 2 Envelope Protein Is Involved in Serotype-Specific Binding to Mosquito but Not Mammalian Cells

2004 ◽  
Vol 78 (1) ◽  
pp. 378-388 ◽  
Author(s):  
Jan-Jong Hung ◽  
Meng-Ti Hsieh ◽  
Ming-Jer Young ◽  
Chuan-Liang Kao ◽  
Chwan-Chuen King ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Envelope Protein ◽  
Mammalian Cells ◽  
Host Cells ◽  
Mammalian Cell Lines ◽  
Loop Region ◽  
Mosquito Cells ◽  
Heparan Sulfates ◽  
Domain Iii

ABSTRACT Dengue virus (DV) is a flavivirus and infects mammalian cells through mosquito vectors. This study investigates the roles of domain III of DV type 2 envelope protein (EIII) in DV binding to the host cell. Recombinant EIII interferes with DV infection to BHK21 and C6/36 cells by blocking dengue virion adsorption to these cells. Inhibition of EIII on BHK21 cells was broad with no serotype specificity; however, inhibition of EIII on C6/36 cells was relatively serotype specific. Soluble heparin completely blocks binding of EIII to BHK21 cells, suggesting that domain III binds mainly to cell surface heparan sulfates. This suggestion is supported by the observation that EIII binds very weakly to gro2C and sog9 mutant mammalian cell lines that lack heparan sulfate. In contrast, heparin does not block binding of EIII to mosquito cells. Furthermore, a synthetic peptide that includes amino acids (aa) 380 to 389 of EIII, IGVEPGQLKL, inhibits binding of EIII to C6/36 but not BHK21 cells. This peptide corresponds to a lateral loop region on domain III of E protein, indicating a possible role of this loop in binding to mosquito cells. In summary, these results suggest that EIII plays an important role in binding of DV type 2 to host cells. In addition, EIII interacts with heparan sulfates when binding to BHK21 cells, and a loop region containing aa 380 to 389 of EIII may participate in DV type 2 binding to C6/36 cells.

scholarly journals Domain-III FG loop of the dengue virus type 2 envelope protein is important for infection of mammalian cells and Aedes aegypti mosquitoes

Virology ◽  
2010 ◽  
Vol 406 (2) ◽  
pp. 328-335 ◽  
Author(s):  
Steven M. Erb ◽  
Siritorn Butrapet ◽  
Kelley J. Moss ◽  
Betty E. Luy ◽  
Thomas Childers ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Dengue Virus ◽  
Virus Type ◽  
Envelope Protein ◽  
Mammalian Cells ◽  
Domain Iii ◽  
Dengue Virus Type 2

scholarly journals DNA Vaccines against Dengue Virus Type 2 Based on Truncate Envelope Protein or Its Domain III

PLoS ONE ◽  
2011 ◽  
Vol 6 (7) ◽  
pp. e20528 ◽  
Author(s):  
Adriana S. Azevedo ◽  
Anna M. Y. Yamamura ◽  
Marcos S. Freire ◽  
Gisela F. Trindade ◽  
Myrna Bonaldo ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Virus Type ◽  
Envelope Protein ◽  
Dna Vaccines ◽  
Domain Iii ◽  
Dengue Virus Type 2

scholarly journals Essential Role of Dengue Virus Envelope Protein N Glycosylation at Asparagine-67 during Viral Propagation

2007 ◽  
Vol 81 (13) ◽  
pp. 7136-7148 ◽  
Author(s):  
Juan A. Mondotte ◽  
Pierre-Yves Lozach ◽  
Ali Amara ◽  
Andrea V. Gamarnik
Keyword(s):  
Dengue Virus ◽  
Envelope Protein ◽  
Mammalian Cells ◽  
Virus Propagation ◽  
Virus Envelope ◽  
Dengue Viruses ◽  
E Protein ◽  
Mosquito Cells ◽  
Glycosylation Sites ◽  
Virus Envelope Protein

ABSTRACT Dengue virus envelope protein (E) contains two N-linked glycosylation sites, at Asn-67 and Asn-153. The glycosylation site at position 153 is conserved in most flaviviruses, while the site at position 67 is thought to be unique for dengue viruses. N-linked oligosaccharide side chains on flavivirus E proteins have been associated with viral morphogenesis, infectivity, and tropism. Here, we examined the relevance of each N-linked glycan on dengue virus E protein by removing each site in the context of infectious viral particles. Dengue viruses lacking Asn-67 were able to infect mammalian cells and translate and replicate the viral genome, but production of new infectious particles was abolished. In addition, dengue viruses lacking Asn-153 in the E showed reduced infectivity. In contrast, ablation of one or both glycosylation sites yielded viruses that replicate and propagate in mosquito cells. Furthermore, we found a differential requirement of N-linked glycans for E secretion in mammalian and mosquito cells. While secretion of E lacking Asn-67 was efficient in mosquito cells, secretion of the same protein expressed in mammalian cells was dramatically impaired. Finally, we found that viruses lacking the carbohydrate at position 67 showed reduced infection of immature dendritic cells, suggesting interaction between this glycan and the lectin DC-SIGN. Overall, our data defined different roles for the two glycans present at the E protein during dengue virus infection, highlighting the involvement of distinct host functions from mammalian and mosquito cells during dengue virus propagation.

scholarly journals Production of a neutralizing antibody against envelope protein of dengue virus type 2 using the linear array epitope technique

2014 ◽  
Vol 95 (10) ◽  
pp. 2155-2165 ◽  
Author(s):  
Peng-Yeh Lai ◽  
Chia-Tse Hsu ◽  
Shao-Hung Wang ◽  
Jin-Ching Lee ◽  
Min-Jen Tseng ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Envelope Protein ◽  
Linear Array ◽  
Polyclonal Antibodies ◽  
Academic Research ◽  
Neutralizing Antibody ◽  
Viral Envelope ◽  
Host Cells ◽  
Viral Envelope Protein ◽  
Domain Iii

Dengue virus (DENV; genus Flavivirus) contains a positive-stranded RNA genome. Binding of DENV to host cells is mediated through domain III of the viral envelope protein. Many therapeutic mAbs against domain III have been generated and characterized because of its high antigenicity. We have previously established a novel PCR method named the linear array epitope (LAE) technique for producing monoclone-like polyclonal antibodies. To prove this method could be utilized to produce antibody against epitopes with low antigenicity, a region of 10 aa (V365NIEAEPPFG374) from domain III of the envelope protein in DENV serotype 2 (DENV2) was selected to design the primers for the LAE technique. A DNA fragment encoding 10 directed repeats of these 10 aa for producing the tandem-repeated peptides was obtained and fused with glutathione S-transferase (GST)-containing vector. This fusion protein (GST-Den EIII10-His6) was purified from Escherichia coli and used as antigen for immunizing rabbits to obtain the polyclonal antibody. Furthermore, the EIII antibody could recognize envelope proteins either ectopically overexpressed or synthesized by DENV2 infection using Western blot and immunofluorescence assays. Most importantly, this antibody was also able to detect DENV2 virions by ELISA, and could block viral entry into BHK-21 cells as shown by immunofluorescence and quantitative real-time PCR assays. Taken together, the LAE technique could be applied successfully for the production of antibodies against antigens with low antigenicity, and shows high potential to produce antibodies with good quality for academic research, diagnosis and even therapeutic applications in the future.

scholarly journals Thioaptamers targeting dengue virus type-2 envelope protein domain III

2014 ◽  
Vol 453 (3) ◽  
pp. 309-315 ◽  
Author(s):  
Sai Hari A. Gandham ◽  
David E. Volk ◽  
Ganesh L.R. Lokesh ◽  
Muniasamy Neerathilingam ◽  
David G. Gorenstein
Keyword(s):  
Dengue Virus ◽  
Virus Type ◽  
Envelope Protein ◽  
Protein Domain ◽  
Domain Iii ◽  
Dengue Virus Type 2

scholarly journals Mutation of the dengue virus type 2 envelope protein heparan sulfate binding sites or the domain III lateral ridge blocks replication in Vero cells prior to membrane fusion

Virology ◽  
2013 ◽  
Vol 441 (2) ◽  
pp. 114-125 ◽  
Author(s):  
John T. Roehrig ◽  
Siritorn Butrapet ◽  
Nathan M. Liss ◽  
Susan L. Bennett ◽  
Betty E. Luy ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Heparan Sulfate ◽  
Membrane Fusion ◽  
Binding Sites ◽  
Envelope Protein ◽  
Vero Cells ◽  
Lateral Ridge ◽  
Domain Iii ◽  
Dengue Virus Type 2

scholarly journals Inhibition of West Nile virus entry by using a recombinant domain III from the envelope glycoprotein

2005 ◽  
Vol 86 (2) ◽  
pp. 405-412 ◽  
Author(s):  
J. J. H. Chu ◽  
R. Rajamanonmani ◽  
J. Li ◽  
R. Bhuvanakantham ◽  
J. Lescar ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Envelope Glycoprotein ◽  
Envelope Protein ◽  
Vero Cells ◽  
Host Cells ◽  
West Nile ◽  
Mosquito Cells ◽  
Outermost Surface ◽  
Host Cell Surface ◽  
Domain Iii

The envelope glycoprotein located at the outermost surface of the flavivirus particle mediates entry of virus into host cells. In this study, the involvement of domain III of West Nile virus (WNV-DIII) envelope protein in binding to host cell surface was investigated. WNV-DIII was first expressed as a recombinant protein and purified after a solubilization and refolding procedure. The refolded WNV-DIII protein displays a content of β-sheets consistent with known homologous structures of other flavivirus envelope DIII, shown by using circular dichroism analysis. Purified recombinant WNV-DIII protein was able to inhibit WNV entry into Vero cells and C6/36 mosquito cells. Recombinant WNV-DIII only partially blocked the entry of dengue-2 (Den 2) virus into Vero cells. However, entry of Den 2 virus into C6/36 was blocked effectively by recombinant WNV-DIII. Murine polyclonal serum produced against recombinant WNV-DIII protein inhibited infection with WNV and to a much lesser extent with Den 2 virus, as demonstrated by plaque neutralization assays. Together these results provided strong evidence that immunoglobulin-like DIII of WNV envelope protein is responsible for binding to receptor on the surface of host cells. The data also suggest that similar attachment molecule(s) or receptor(s) were used by WNV and Den 2 virus for entry into C6/36 mosquito cells.

scholarly journals Evolutionary Selection of the Nuclear Localization Signal in the Viral Nucleoprotein Leads to Host Adaptation of the Genus Orthobornavirus

Viruses ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 1291
Author(s):  
Ryo Komorizono ◽  
Yukiko Sassa ◽  
Masayuki Horie ◽  
Akiko Makino ◽  
Keizo Tomonaga
Keyword(s):  
Host Range ◽  
Nuclear Localization ◽  
Nuclear Localization Signal ◽  
Mammalian Cells ◽  
Matrix Protein ◽  
Host Adaptation ◽  
Host Cells ◽  
Viral Glycoprotein ◽  
Mammalian Cell Lines ◽  
Localization Signal

Adaptation of the viral life cycle to host cells is necessary for efficient viral infection and replication. This evolutionary process has contributed to the mechanism for determining the host range of viruses. Orthobornaviruses, members of the family Bornaviridae, are non-segmented, negative-strand RNA viruses, and several genotypes have been isolated from different vertebrate species. Previous studies revealed that some genotypes isolated from avian species can replicate in mammalian cell lines, suggesting the zoonotic potential of avian orthobornaviruses. However, the mechanism by which the host specificity of orthobornaviruses is determined has not yet been identified. In this study, we found that the infectivity of orthobornaviruses is not determined at the viral entry step, mediated by the viral glycoprotein and matrix protein. Furthermore, we demonstrated that the nuclear localization signal (NLS) sequence in the viral nucleoprotein (N) has evolved under natural selection and determines the host-specific viral polymerase activity. A chimeric mammalian orthobornavirus, which has the NLS sequence of avian orthobornavirus N, exhibited a reduced propagation efficiency in mammalian cells. Our findings indicated that nuclear transport of the viral N is a determinant of the host range of orthobornaviruses, providing insights into the evolution and host adaptation of orthobornaviruses.

Sign in / Sign up

Export Citation Format

Share Document