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Cell-based analysis of Chikungunya virus membrane fusion using baculovirus-expression vectors.

Authors :
Kuo SC
Chen YJ
Wang YM
Kuo MD
Jinn TR
Chen WS
Chang YC
Tung KL
Wu TY
Lo SJ
Source :
Journal of virological methods [J Virol Methods] 2011 Aug; Vol. 175 (2), pp. 206-15. Date of Electronic Publication: 2011 May 17.
Publication Year :
2011

Abstract

Chikungunya virus infection has emerged in many countries over the past decade. There are no effective drugs for controlling the disease. To develop cell-based system for screening anti-virus drugs, a bi-cistronic baculovirus expression system was utilized to co-express viral structural proteins C (capsid), E2 and E1 and the enhanced green fluorescence protein (EGFP) in Spodoptera frugiperda insect cells (Sf21). The EGFP-positive Sf21 cells fused with each other and with uninfected cells to form a syncytium, allowing characterization of cholesterol and low pH requirements for syncytium formation. Western blot analysis showed three structural proteins were expressed in baculovirus infected cells. The structural proteins of Chikungunya virus that is required for cell fusion was determined with various recombinant baculoviruses bearing different lengths of the viral structural protein genes. Protein E1 was required for cell fusion and indicating that Chikungunya viral membrane fusion was a class II membrane fusion. It was also demonstrated that the heterologous expression of alphavirus monomeric E1 can induce insect cell fusions. Furthermore, this cell-based system provides a model for studying class II viral membrane fusion.<br /> (Copyright © 2011 Elsevier B.V. All rights reserved.)

Details

Language :
English
ISSN :
1879-0984
Volume :
175
Issue :
2
Database :
MEDLINE
Journal :
Journal of virological methods
Publication Type :
Academic Journal
Accession number :
21619896
Full Text :
https://doi.org/10.1016/j.jviromet.2011.05.015