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Journal of Virology, February 2001, p. 1427-1436, Vol. 75, No. 3
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.3.1427-1436.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Role of the UL25 Gene Product in Packaging DNA into the Herpes Simplex Virus Capsid: Location of UL25 Product in the Capsid and Demonstration that It Binds DNA

Masahiro Ogasawara,* Tatsuo Suzutani, Itsuro Yoshida, and Masanobu Azuma

Department of Microbiology, Asahikawa Medical College, 2-1-1-1, Midorigaoka-Higashi, Asahikawa 078-8510, Japan

Received 3 July 2000/Accepted 6 November 2000

Recent studies have suggested that the herpes simplex type 1 (HSV-1) UL25 gene product, a minor capsid protein, is required for encapsidation but not cleavage of replicated viral DNA. This study set out to investigate the potential interactions of UL25 protein with other virus proteins and determine what properties it has for playing a role in DNA encapsidation. The UL25 protein is found in 42 ± 17 copies per B capsid and is present in both pentons and hexons. We introduced green fluorescent protein (GFP) as a fluorescent tag into the N terminus of UL25 protein to identify its location in HSV-1-infected cells and demonstrated the relocation of UL25 protein from the cytoplasm into the nucleus at the late stage of HSV-1 infection. To clarify the cause of this relocation, we analyzed the interactions of UL25 protein with other virus proteins. The UL25 protein associates with VP5 and VP19C of virus capsids, especially of the penton structures, and the association with VP19C causes its relocation into the nucleus. Gel mobility shift analysis shows that UL25 protein has the potential to bind DNA. Moreover, the amino-terminal one-third of the UL25 protein is particularly important in DNA binding and forms a homo-oligomer. In conclusion, the UL25 gene product forms a tight connection with the capsid being linked with VP5 and VP19C, and it may play a role in anchoring the genomic DNA.

* Corresponding author. Mailing address: Department of Microbiology, Asahikawa Medical College, 2-1-1-1, Midorigaoka-Higashi, Asahikawa 078-8510, Japan. Phone: 81-166-68-2393. Fax: 81-166-68-2399. E-mail: kazuyou{at}asahikawa-med.ac.jp.

Journal of Virology, February 2001, p. 1427-1436, Vol. 75, No. 3
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.3.1427-1436.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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