Identification of a DNA encapsidation sequence for human polyomavirus pseudovirion formation
Wei-Chih Ou
Institute of Molecular Biology, National Chung Cheng University, Chia-Yi, Taiwan
Institute of Life Science, National Tsing Hua University, Hsinchu, Taiwan
Search for more papers by this authorTzong-Hsiung Hseu
Institute of Life Science, National Tsing Hua University, Hsinchu, Taiwan
Search for more papers by this authorMeilin Wang
Department of Microbiology and Immunology, Chung Shan Medical and Dental College, Taichung, Taiwan
Search for more papers by this authorHan Chang
Department of Pathology, Chung Shan Hospital, Taichung, Taiwan
Search for more papers by this authorCorresponding Author
Deching Chang
Institute of Molecular Biology, National Chung Cheng University, Chia-Yi, Taiwan
Department of Microbiology and Immunology, Chung Shan Medical and Dental College, No.110, Section 1, Chien Kou N. Road, Taichung 402, Taiwan.===Search for more papers by this authorWei-Chih Ou
Institute of Molecular Biology, National Chung Cheng University, Chia-Yi, Taiwan
Institute of Life Science, National Tsing Hua University, Hsinchu, Taiwan
Search for more papers by this authorTzong-Hsiung Hseu
Institute of Life Science, National Tsing Hua University, Hsinchu, Taiwan
Search for more papers by this authorMeilin Wang
Department of Microbiology and Immunology, Chung Shan Medical and Dental College, Taichung, Taiwan
Search for more papers by this authorHan Chang
Department of Pathology, Chung Shan Hospital, Taichung, Taiwan
Search for more papers by this authorCorresponding Author
Deching Chang
Institute of Molecular Biology, National Chung Cheng University, Chia-Yi, Taiwan
Department of Microbiology and Immunology, Chung Shan Medical and Dental College, No.110, Section 1, Chien Kou N. Road, Taichung 402, Taiwan.===Search for more papers by this authorAbstract
Human polyomavirus is a naked capsid virus containing a closed circular double-stranded DNA genome. The mechanism of DNA encapsidation for the viral progeny formation is not fully understood. In this study, DNA encapsidation domain of the major capsid protein, VP1, of the human polyomavirus JCV was investigated. When the first 12 amino acids were deleted, the E. coli expressed VP1 (ΔN12VP1) failed to encapsidate the host DNA although the integrity of the capsid-like structure was maintained. In addition, capsid-like particles of ΔN12VP1 did not package exogenous DNA in vitro, which is in contrast to that of the full-length VP1 protein. These findings suggest that the N-terminal of the first 12 amino acids of VP1 were responsible for DNA encapsidation. The importance of amino acids in the DNA encapsidation domain was determined further using site-directed mutagenesis. All of the positively charged amino acids at the N-terminal region of VP1 were essential for DNA encapsidation. The results indicate that the N-terminal region of the human polyomavirus major capsid protein VP1 may be involved in viral genome encapsidation during progeny maturation. J. Med. Virol. 64:366–373, 2001. © 2001 Wiley-Liss, Inc.
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