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Journal of Virology, November 2003, p. 12357-12362, Vol. 77, No. 22
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.22.12357-12362.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

RNA Helicase Domain of Tobamovirus Replicase Executes Cell-to-Cell Movement Possibly through Collaboration with Its Nonconserved Region

Kyotaro Hirashima1,2 and Yuichiro Watanabe1*

Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Meguro-ku, Tokyo 153-8902,1 R&D Group, ASPEX Division, Asahi Glass Co., Ltd., Kanagawa-ku, Yokohama-shi, Kanagawa 221-8755, Japan2

Received 19 May 2003/ Accepted 1 August 2003

UR-hel, a chimeric virus obtained by replacement of the RNA helicase domain of tobacco mosaic virus (TMV)-U1 replicase with that from the TMV-R strain, could replicate similarly to TMV-U1 in protoplasts but could not move from cell to cell (K. Hirashima and Y. Watanabe, J. Virol. 75:8831-8836, 2001). It was suggested that TMV recruited both the movement protein (MP) and replicase for cell-to-cell movement by unknown mechanisms. Here, we found that a recombinant, UR-hel/V, in which the nonconserved region was derived from TMV-R in addition to the RNA helicase domain of replicase, could move from cell to cell. We also analyzed revertants isolated from UR-hel, which recovered cell-to-cell movement by their own abilities. We found amino acid substitutions responsible for phenotypic reversion only in the nonconserved region and/or RNA helicase domain but never in MP. Together, these data show that both the nonconserved region and the RNA helicase domain of replicase are involved in cell-to-cell movement. The RNA helicase domain of tobamovirus replicase possibly does not interact directly with MP but interacts with its nonconserved region to execute cell-to-cell movement.

* Corresponding author. Mailing address: Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Komaba 3-8-1, Meguro-ku, Tokyo 153-8902, Japan. Phone: 81-3-5454-6776. Fax: 81-3-5454-6776. E-mail: solan{at}bio.c.u-tokyo.ac.jp.

Journal of Virology, November 2003, p. 12357-12362, Vol. 77, No. 22
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.22.12357-12362.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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