Restriction of human immunodeficiency virus type 1 Rev function in murine A9 cells involves the Rev C-terminal domain.
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.
Marques
Sandra M P
SM
Department of Biochemistry and Molecular Biology, The George Washington University, Washington, DC 20037, USA.
Veyrune
Jean-Luc
JL
Shukla
Ram R
RR
Kumar
Ajit
A
D000818:Animals; D002460:Cell Line; D005347:Fibroblasts / Q000821:virology*; D015967:Gene Expression Regulation, Viral; D015683:Gene Products, gag / Q000235:genetics / Q000378:metabolism; D015694:Gene Products, rev / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D016370:Gene Products, rex / Q000378:metabolism; D049452:Green Fluorescent Proteins; D015497:HIV-1 / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D054321:rev Gene Products, Human Immunodeficiency Virus
D015683:Gene Products, gag; D015694:Gene Products, rev; D016370:Gene Products, rex; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D054321:rev Gene Products, Human Immunodeficiency Virus; D049452:Green Fluorescent Proteins