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Journal of Virology, April 2004, p. 3889-3896, Vol. 78, No. 8
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.8.3889-3896.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Poly(A)- and Primer-Independent RNA Polymerase of Norovirus

Shuetsu Fukushi,1,{dagger} Shigeyuki Kojima,1 Reiko Takai,1 Fuminori B. Hoshino,1 Tomoichiro Oka,2 Naokazu Takeda,2 Kazuhiko Katayama,2 and Tsutomu Kageyama1*

R&D Center, BioMedical Laboratories, Kawagoe, Saitama 350-1101,1 Department of Virology II, National Institute of Infectious Diseases, Musashi-Murayama, Tokyo 208-0011, Japan2

Received 27 August 2003/ Accepted 27 December 2003

Replication of positive-strand caliciviruses is mediated by a virus-encoded RNA-dependent RNA polymerase (RdRp). To study the replication of Norovirus (NV), a member of the family Caliciviridae, we used a recombinant baculovirus system to express an enzymatically active RdRp protein from the 3D region of the NV genome and defined conditions for optimum enzymatic activity. Using an RNA template from the NV 3' genomic region, we observed similar levels of enzymatic activity in assays with and without a poly(A) tail. RdRp activity was not significantly affected by the addition of an RNA primer to the reaction mixture. Thus, the NV RdRp exhibited primer- and poly(A)-independent RNA polymerase activity. While the RdRp inhibitor phosphonoacetic acid inhibited NV RdRp activity, another gliotoxin did not. The active recombinant NV RdRp will be of benefit to studies of NV replication and will facilitate the development of specific inhibitors of NV proliferation.

* Corresponding author. Mailing address: R&D Center, BioMedical Laboratories, Matoba 1361-1, Kawagoe, Saitama 350-1101, Japan. Phone: (81)49-232-0440. Fax: (81)49-232-5480. E-mail: tkage{at}alk.co.jp.

{dagger} Present address: Department of Virology I, National Institute of Infectious Diseases, Tokyo 208-0011, Japan.

Journal of Virology, April 2004, p. 3889-3896, Vol. 78, No. 8
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.8.3889-3896.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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