Autor(es): Kuo Szu-Cheng, Chen Ying-Ju, Wang Yu-Ming, Kuo Ming-Der, Jinn Tzyy-Rong, Chen Wen-Shuo, Chang Yen-Chung, Tung Kuo-Lun, Wu Tzong-Yuan, Lo Szecheng J

Resumo: 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 - E1 - the enhanced green fluorescence protein (EGFP) in Spodoptera frugiperda insect cells (Sf21). The EGFP-positive Sf21 cells fused with each other - with uninfected cells to form a syncytium, allowing characterization of cholesterol - 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 - 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.

Palavras-Chave: Baculovirus; Chikungunya virus; Internal ribosome entry site; Syncytium; Viral membrane fusion

Imprenta: Journal of Virological Methods, v. 175, n. 2, p. 206-215, 2011

Identificador do objeto digital: 10.1016/j.jviromet.2011.05.015

Descritores: Chikungunya virus - Biosynthesis ; Chikungunya virus - Cell ; Chikungunya virus - Genome ; Chikungunya virus - Pathogenesis ; Chikungunya virus - Protein synthesis ; Chikungunya virus - Proteins ; Chikungunya Virus - Virus

Data de publicação: 2011

INDICADORES

ACESSO