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Journal of Virology, July 2009, p. 6624-6630, Vol. 83, No. 13
0022-538X/09/$08.00+0     doi:10.1128/JVI.02385-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Different Subcellular Localization of Theiler's Murine Encephalomyelitis Virus Leader Proteins of GDVII and DA Strains in BHK-21 Cells{triangledown}

Naoko Taniura, Mineki Saito, Takako Okuwa, Kousuke Saito, and Yoshiro Ohara*

Department of Microbiology, Kanazawa Medical University School of Medicine, Uchinada, Ishikawa 920-0293, Japan

Received 17 November 2008/ Accepted 13 April 2009

The highly virulent GDVII strain of Theiler's murine encephalomyelitis virus causes acute and fatal encephalomyelitis, whereas the DA strain causes mild encephalomyelitis followed by a chronic inflammatory demyelinating disease with virus persistence. The differences in the amino acid sequences of the leader protein (L) of the DA and GDVII strains are greater than those for any other viral protein. We examined the subcellular distribution of DA L and GDVII L tagged with the FLAG epitope in BHK-21 cells. Wild-type GDVII L was localized predominantly in the cytoplasm, whereas wild-type DA L showed a nucleocytoplasmic distribution. A series of the L mutant experiments demonstrated that the zinc finger domain, acidic domain, and C-terminal region of L were necessary for the nuclear accumulation of DA L. A GDVII L mutant with a deletion of the serine/threonine (S/T)-rich domain showed a nucleocytoplasmic distribution, in contrast to the predominant cytoplasmic distribution of wild-type GDVII L. A chimeric DA/GDVII L, D/G, which encodes the N region of DA L including the zinc finger domain and acidic domain, followed by the GDVII L sequence including the S/T-rich domain, was distributed exclusively throughout the cytoplasm but not in the nucleus, as observed with wild-type GDVII L. Another chimeric L, G/D (which is the converse of the D/G construct), accumulated in the nucleus as well as the cytoplasm, as was observed for wild-type DA L. The findings suggest that the differential distribution of DA L and GDVII L is determined primarily by the S/T-rich domain. The S/T-rich domain may be important for the viral activity through the regulation of the subcellular distribution of L.

* Corresponding author. Mailing address: Department of Microbiology, Kanazawa Medical University School of Medicine, Uchinada, Ishikawa 920-0293, Japan. Phone: 81-76-218-8096. Fax: 81-76-286-3961. E-mail: ohara{at}kanazawa-med.ac.jp

{triangledown} Published ahead of print on 22 April 2009.

Journal of Virology, July 2009, p. 6624-6630, Vol. 83, No. 13
0022-538X/09/$08.00+0     doi:10.1128/JVI.02385-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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