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Journal of Virology, March 2001, p. 2331-2336, Vol. 75, No. 5
Division of Molecular Genetics, National
Institute of Infectious Diseases, Shinjuku-ku, Tokyo
162-8640,1 and Department of Obstetrics
and Gynecology, Faculty of Medicine, University of Tokyo,
Bunkyo-ku, Tokyo 113-0033,2 Japan
Received 24 August 2000/Accepted 29 November 2000
The first step of papillomavirus infection is believed to be
binding of major capsid protein L1 to the cell surface without involvement of minor capsid protein L2, but the viral infectivity can
be neutralized either by anti-L1 or anti-L2 antibody. To understand the
role of L2 in human papillomavirus (HPV) infection, we examined a
segment of HPV type 16 (HPV16) L2, which contains a neutralization epitope common to HPV6, for its involvement in adsorption and penetration of the capsids. Preincubation of monkey COS-1 cells with a
synthetic peptide having amino acids (aa) 108 to 120 of HPV16 L2
reduced the susceptibility of COS-1 cells to infection with HPV16
pseudovirions. Confocal microscopy showed that the green fluorescence
protein (GFP) fused with the L2 peptide was found to bind to the
surface of a HeLa cell, an HPV18-positive human cancer cell line, at
4°C and to enter the cytoplasm after subsequent incubation at 37°C.
Flow cytometry showed that fused GFP did not bind to HeLa cells that
had been treated with trypsin. Besides COS-1 and HeLa cells, some human
and rodent cell lines were detected by flow cytometry to be susceptible
to binding with fused GFP, showing a tendency of epithelial cells
toward higher susceptibility. Substitutions at aa 108 to 111 inhibited
fused GFP from binding to HeLa cells and reduced the infectivity in COS-1 cells of the in vitro-constructed pseudovirions. The results suggest that L2 plays an important role in enhancing HPV infection through interaction between the N-terminal region and a cellular surface protein, facilitating penetration of the virions and
determining part of the tropism of HPVs.
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.5.2331-2336.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Human Papillomavirus Type 16 Minor Capsid Protein L2 N-Terminal
Region Containing a Common Neutralization Epitope Binds to the Cell
Surface and Enters the Cytoplasm
*
Corresponding author. Mailing address: Division of
Molecular Genetics, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan. Phone: (81)-3-5285-1111,
ext. 2524. Fax: (81)-3-5285-1166. E-mail:
kanda{at}nih.go.jp.
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