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Journal of Virology, October 1999, p. 8112-8119, Vol. 73, No. 10
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Human T-Cell Leukemia Virus Type 2 Rex Protein Increases Stability and Promotes Nuclear to Cytoplasmic Transport of gag/pol and env RNAs

Koichi Kusuhara,1,dagger Matthew Anderson,1 Sherrie M. Pettiford,1 and Patrick L. Green2,*

Department of Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-2363,1 and Department of Veterinary Biosciences and Molecular Virology, Immunology, and Medical Genetics, Center for Retrovirus Research, and Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio 43210-10932

Received 13 April 1999/Accepted 6 July 1999

The human T-cell leukemia virus (HTLV) Rex protein is essential for efficient expression of the viral structural and enzymatic gene products. In this study, we assessed the role of the HTLV-2 rex gene in viral RNA expression and Gag protein production. Following transfection of human JM4 T cells with wild-type and rex mutant full-length proviral constructs, PCR was used for semiquantitative analysis of specific viral RNA transcripts. In the presence of Rex, the total amount of steady-state viral RNA was increased fourfold. Rex significantly up-regulated the level of incompletely spliced RNAs by increasing RNA stability and was associated with a twofold down-regulation of the completely spliced tax/rex RNA. PCR analysis of subcellular RNA fractions, isolated from transfected cells, indicated that the level of gag/pol and env cytoplasmic RNAs were increased 7- to 9-fold in the presence of Rex, whereas Gag protein production was increased 130-fold. These data indicate that HTLV-2 Rex increases the stability and promotes nucleus-to-cytoplasm transport of the incompletely spliced viral RNAs, ultimately resulting in increased structural protein production. Moreover, this model system provides a sensitive approach to further characterize HTLV gene expression from full-length proviral clones following transfection of human T cells.

* Corresponding author. Mailing address: Department of Veterinary Biosciences, The Ohio State University, 1925 Coffey Rd., Columbus, OH 43210-1093. Phone: (614) 688-4899. Fax: (614) 292-6473. E-mail: green.466{at}osu.edu.

dagger Present address: Department of Pediatrics, Faculty of Medicine, Kyushu University, Higashi-ku, Fukuoka 812-8582, Japan.

Journal of Virology, October 1999, p. 8112-8119, Vol. 73, No. 10
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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