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Journal of Virology, March 2003, p. 3084-3090, Vol. 77, No. 5
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.5.3084-3090.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Restriction of Human Immunodeficiency Virus Type 1 Rev Function in Murine A9 Cells Involves the Rev C-Terminal Domain

Sandra M. P. Marques,1,2 Jean-Luc Veyrune,2,{dagger} Ram R. Shukla,1,2,{ddagger} and Ajit Kumar1,2*

Graduate Program in Genetics,1 Department of Biochemistry and Molecular Biology, The George Washington University, Washington, D.C. 200372

Received 1 July 2002/ Accepted 3 December 2002

The human immunodeficiency virus type 1 (HIV-1) Rev and human T-cell leukemia virus type 1 (HTLV-1) Rex proteins are essential for the expression of viral structural proteins and productive infection. Both contain a nuclear export signal (NES) in their C-terminal domain and a nuclear localization signal (NLS) in their N-terminal domain. The NES and NLS are necessary for shuttling between nucleus and cytoplasm and are therefore indispensable for the transport of unspliced and singly spliced viral transcripts. HIV-1 Rev function is restricted in A9 cells, a murine fibroblast cell line, whereas HTLV-1 Rex is functional in these cells. Immunofluorescence studies with RevGFP fusion protein demonstrate normal import and export of Rev in A9 cells. To ascertain which domains of Rev are necessary for the restriction of Rev function in A9 cells, we studied a chimeric construct in which the NES domain of Rev was exchanged with Rex C-terminal amino acids 79 to 95, the Rev1-79/Rex79-95 chimera, which restored Rev function in A9 cells. In addition, overexpression of a truncated Rev containing the Rev C-terminal domain in the presence of wild-type Rev, led to restoration of Rev function in A9 cells. These results suggest that the C-terminal domain of HIV-1 Rev plays an important role in restricting Rev function in murine cells.

* Corresponding author. Mailing address: Department of Biochemistry and Molecular Biology, The George Washington University, Washington, DC 20037. Phone: (202) 994-2919. Fax: (202) 994-8974. E-mail: akumar{at}gwu.edu.

{dagger} Present address: Unité de Therapie Cellulaire, Hôpital Saint Eloi, Montpellier, France.

{ddagger} Present address: United States Patent and Trademark Office, Crystal City, Va.

Journal of Virology, March 2003, p. 3084-3090, Vol. 77, No. 5
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.5.3084-3090.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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