Journal of General Virology, Vol 78, 1435-1439, Copyright © 1997 by Society for General Microbiology

ARTICLES

Self-assembly of the JC virus major capsid protein, VP1, expressed in insect cells

D Chang, CY Fung, WC Ou, PC Chao, SY Li, M Wang, YL Huang, TY Tzeng and RT Tsai
Department of Microbiology, Chung Shan Medical and Dental College, Taichung, Taiwan, Republic of China. dch@mercury.csmc.edu.tw

The major capsid protein of human polyomavirus JC virus, VP1, has been cloned into a baculovirus genome and expressed in insect cells. The VP1 protein was expressed in the cytoplasm and transported into the nucleus. It was then purified by a sucrose cushion and CsCI density gradient centrifugation to near homogeneity. Electron microscopy showed that isolated recombinant VP1 protein self-assembled into a capsid-like structure similar to the natural empty capsid. Both chelator (EDTA) and reducing agent (DTT) are required to disrupt the capsid structure into the pentameric capsomeres, as demonstrated by haemagglutination assay and electron microscopy. These results suggest that JC virus VP1 can be transported into the nucleus and self-assembled to form capsid-like particles without the involvement of the viral minor capsid proteins, VP2 and VP3. In addition, metal ions and disulphide bonds appear to be important in maintaining the integrity of the viral capsid structure.

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