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J Virol, May 1998, p. 3935-3943, Vol. 72, No. 5
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

The Transmembrane Domains of Sindbis Virus Envelope Glycoproteins Induce Cell Death

Andrew K. Joe, Hsin Hsin Foo, Linda Kleeman, and Beth Levine*

Department of Medicine, Columbia University College of Physicians and Surgeons, New York, New York 10032

Received 29 August 1997/Accepted 12 January 1998

Sindbis virus, the prototype alphavirus, kills cells by inducing apoptosis. To investigate potential mechanisms by which Sindbis virus induces apoptosis, we examined whether specific viral gene products were able to induce cell death. Genes encoding the three structural proteins---capsid, the precursor E1 (6K plus E1), and the precursor E2 (P62 or E3 plus E2)---were cotransfected with a beta -galactosidase reporter plasmid in transient-transfection assays in rat prostate adenocarcinoma AT3 cells. Cell death, as determined by measuring the loss of blue cells, was observed in AT3 cells transfected with 6K plus E1 and with P62 but not in cells transfected with capsid. Deletion mutagenesis of P62 indicated that large regions of the cytoplasmic domain and extracellular domain were not essential for the induction of cell death. However, constructs containing the minimal E3 signal sequence fused to the E2 transmembrane domain and the minimal E3 signal sequence fused to the E1 transmembrane domain induced death as efficiently as full-length P62 and 6K plus E1, whereas no cell death was observed after transfection with a control construct containing the E3 signal sequence linked to the transmembrane domain of murine CD4. These data demonstrate that intracellular expression of the transmembrane domains of the Sindbis virus envelope glycoproteins can kill AT3 cells.

* Corresponding author. Mailing address: Department of Medicine, Columbia University College of Physicians and Surgeons, 630 W. 168th St., New York, NY 10032. Phone: (212) 305-7312. Fax: (212) 305-7290. E-mail: Levine{at}cuccfa.ccc.columbia.edu.

J Virol, May 1998, p. 3935-3943, Vol. 72, No. 5
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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