Chimeric flavivirus between Binjari virus and West Nile (Kunjin) virus

Most known flaviviruses, including West Nile virus (WNV), are maintained in natural transmission cycles between hematophagous arthropods and vertebrate hosts. Other flaviviruses such as Modoc virus (MODV) and Culex flavivirus (CxFV) have host ranges restricted to vertebrates and insects, respectively. The genetic elements that modulate the differential host ranges and transmission cycles of these viruses have not been identified. Fusion polymerase chain reaction (PCR) was used to replace the capsid (C), premembrane (prM) and envelope (E) genes and the prM-E genes of a full-length MODV infectious cDNA clone with the corresponding regions of WNV and CxFV. Fusion products were directly transfected into baby hamster kidney-derived cells that stably express T7 RNA polymerase. At 4 days post-transfection, aliquots of each supernatant were inoculated onto vertebrate (BHK-21 and Vero) and mosquito (C6/36) cells which were then assayed for evidence of viral infection by reverse transcription-PCR, Western blot and plaque assay. Chimeric virus was recovered in cells transfected with the fusion product containing the prM-E genes of WNV. The virus could infect vertebrate but not mosquito cells. The in vitro replication kinetics and yields of the chimeric virus were similar to MODV but the chimeric virus produced larger plaques. Chimeric virus was not recovered in cells transfected with any of the other fusion products. Our data indicate that genetic elements outside of the prM-E gene region of MODV condition its vertebrate-specific phenotype.

Vero cell

Flavivirus

Chimeric gene

Recombinant virus

10.1186/1743-422x-11-150

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Citations (12)

Interaction of West Nile and Kunjin Viruses with Cellular Components During Morphogenesis

Current topics in microbiology and immunology (2002)

Mah Lee Ng Justin Jang Hann Chu

West Nile virus

Replication

10.1007/978-3-642-59403-8_17

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Citations (12)

Identification and characterization of cell-adapted mutations in West Nile Virus and replicons

Shannan L. Rossi

West Nile virus

Identification

Replicon

Source

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Citations (0)

Yellow fever vector live-virus vaccines: West Nile virus vaccine development

Trends in Molecular Medicine (2001)

Juan Arroyo Charles A. Miller John Catalan Thomas P. Monath

Flavivirus

Attenuated vaccine

Dengue vaccine

West Nile virus

10.1016/s1471-4914(01)02048-2

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Citations (59)

Chimeric flavivirus between Binjari virus and Murray Valley encephalitis virus

Joshua M. Hardy H.V. Venugopal Natalee D. Newton Daniel Watterson Fasséli Coulibaly

Flavivirus

10.2210/pdb7kvb/pdb

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Citations (0)

Development and characterization of non-glycosylated E and NS1 mutant viruses as a potential candidate vaccine for West Nile virus

Vaccine (2009)

Melissa C. Whiteman Li Li Jason A. Wicker Richard M. Kinney Claire Y.‐H. Huang

Flavivirus

Viremia

Serial passage

10.1016/j.vaccine.2009.10.112

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Citations (75)

Biological Significance of Glycosylation of the Envelope Protein of Kunjin Virus

Annals of the New York Academy of Sciences (2001)

J. Scherret J. S. Mackenzie Alexander A. Khromykh Roy A. Hall

Parasitology

Clinical Microbiology

Annals

Tropical Medicine

Memphis

10.1111/j.1749-6632.2001.tb02719.x

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Citations (60)

Kunjin virus and Kunjin encephalitis

J. S. Mackenzie

Source

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Citations (0)

West Nile Virus Capsid and viral RNA interaction

International Journal of Infectious Diseases (2008)

Yuen Kuen Cheong Mah Lee Ng

Background and Aim: The process of assembly of the West Nile (WN) virus presents itself as an attractive anti-viral target. This involves both oligomerization of the capsid (C) protein and packaging of the viral RNA. The RNA binding properties of the C protein is poorly understood. This study aims to characterize the C and RNA interaction. Method: Co-localization study was performed by transfecting in vitro transcribed WN virus RNA and C protein clones into BHK cells and visualized under fluorescence microscopy. RNA binding properties of C protein were further investigated with a Northwestern Blot assay and RNA pull-down assay. Synthesized C protein peptides were used to map out the RNA binding regions on C protein. In addition, C protein immuno-purified from BHK cells were used to investigate the effect of phosphorylation of C protein on its RNA binding properties. Results: RNA and C protein have failed to show co-localization in BHK cells by immuno-fluorescence but interactions were observed at the molecular level. It showed that the first 465 and last 693 nucleotides of the WN virus RNA had specific affinity for the full length C protein. In addition, the amino- and carboxy-terminal of the C protein were shown to bind to the virus RNA. It was also found that the C protein had affinity for viral anti-sense RNA. Phosphorylated peptides of C protein and C protein expressed in BHK cells show attenuated binding to viral RNA. Conclusion: C protein interaction with anti-sense indicates that its interaction with viral sense RNA may not be specific. Phosphorylation of C protein could play a role in regulating C and RNA interaction and allow time for viral assembly. Understanding the interaction of C protein with viral RNA and role of phosphorylation in nucleocapsid assembly could help develop anti-viral strategies aimed at disrupting viral assembly.

Viral protein

Viral structural protein

Nuclease protection assay

10.1016/j.ijid.2008.05.787

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