Essential elements of the capsid protein for self-assembly into empty virus-like particles of hepatitis E virus.
Hepatitis E virus (HEV) is a noncultivable virus that causes acute liver failure in humans. The virus's major capsid protein is encoded by an open reading frame 2 (ORF2) gene. When the recombinant protein consisting of amino acid (aa) residues 112 to 660 of ORF2 is expressed with a recombinant baculovirus, the protein self-assembles into virus-like particles (VLPs) (T.-C. Li, Y. Yamakawa, K. Suzuki, M. Tatsumi, M. A. Razak, T. Uchida, N. Takeda, and T. Miyamura, J. Virol. 71:7207-7213, 1997). VLPs can be found in the culture medium of infected Tn5 cells but not in that of Sf9 cells, and the major VLPs have lost the C-terminal 52 aa. To investigate the protein requirement for HEV VLP formation, we prepared 14 baculovirus recombinants to express the capsid proteins truncated at the N terminus, the C terminus, or both. The capsid protein consisting of aa residues 112 to 608 formed VLPs in Sf9 cells, suggesting that particle formation is dependent on the modification process of the ORF2 protein. In the present study, electron cryomicroscopy and image processing of VLPs produced in Sf9 and Tn5 cells indicated that they possess the same configurations and structures. Empty VLPs were found in both Tn5 and Sf9 cells infected with the recombinant containing an N-terminal truncation up to aa residue 125 and C-terminal to aa residue 601, demonstrating that the aa residues 126 to 601 are the essential elements required for the initiation of VLP assembly. The recombinant HEV VLPs are potential mucosal vaccine carrier vehicles for the presentation of foreign antigenic epitopes and may also serve as vectors for the delivery of genes to mucosal tissue for DNA vaccination and gene therapy. The results of the present study provide useful information for constructing recombinant HEV VLPs having novel functions.
Li
Tian-Cheng
TC
Department of Virology II, National Institute of Infectious Diseases, Musashi-Murayama, Tokyo, Japan. litc@nih.go.jp
Takeda
Naokazu
N
Miyamura
Tatsuo
T
Matsuura
Yoshiharu
Y
Wang
Joseph C Y
JC
Engvall
Helena
H
Hammar
Lena
L
Xing
Li
L
Cheng
R Holland
RH
D000818:Animals; D016367:Baculoviridae / Q000378:metabolism; D036022:Capsid Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D016752:Hepatitis E virus / Q000737:chemistry* / Q000502:physiology; D008958:Models, Molecular; D016366:Open Reading Frames; D011994:Recombinant Proteins / Q000096:biosynthesis; D019065:Virus Assembly
D036022:Capsid Proteins; D011994:Recombinant Proteins