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Journal of Virology, June 2002, p. 5422-5434, Vol. 76, No. 11
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.11.5422-5434.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Transduction of Interphase Cells by Avian Sarcoma Virus

Richard A. Katz,1* James G. Greger,1,2 Kristen Darby,1,{dagger} Pamela Boimel,1,{ddagger} Glenn F. Rall,1 and Anna Marie Skalka1,2

Institute for Cancer Research, Fox Chase Cancer Center,1 Graduate Group in Cell and Molecular Biology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania2

Received 13 November 2001/ Accepted 20 February 2002

It has been generally believed that oncoretroviruses are dependent on mitosis for efficient nuclear entry of viral DNA. We previously identified a nuclear localization signal in the integrase protein of an oncoretrovirus, avian sarcoma virus (ASV), suggesting an active import mechanism for the integrase-DNA complex (G. Kukolj, R. A. Katz, and A. M. Skalka, Gene 223:157-163, 1998). Here, we have evaluated the requirement for mitosis in nuclear import and integration of ASV DNA. Using a modified ASV encoding a murine leukemia virus amphotropic env gene and a green fluorescent protein (GFP) reporter gene, DNA nuclear import was measured in cell cycle-arrested avian (DF-1) as well as human (HeLa) and mouse cells. The results showed efficient accumulation of nuclear forms of ASV DNA in {gamma}-irradiation-arrested cells. Efficient transduction of a GFP reporter gene was also observed after infection of cells that were arrested with {gamma}-irradiation, mitomycin C, nocodazole, or aphidicolin, confirming that nuclear import and integration of ASV DNA can occur in the absence of mitosis. By monitoring GFP expression in individual cells, we also obtained evidence for nuclear import of viral DNA during interphase in cycling cells. Lastly, we observed that ASV can transduce postmitotic mouse neurons. These results support an active nuclear import mechanism for the oncoretrovirus ASV and suggest that this mechanism can operate in both nondividing and dividing cells.

* Corresponding author. Mailing address: Institute for Cancer Research, Fox Chase Cancer Center, 7701 Burholme Ave., Philadelphia, PA 19111. Phone: (215) 728-3668. Fax: (215) 728-2778. E-mail: R_Katz{at}fccc.edu.

{dagger} Present address: Merck & Co., Inc., West Point, PA 19486.

{ddagger} Present address: Drexel University, Philadelphia, PA 19104.

Journal of Virology, June 2002, p. 5422-5434, Vol. 76, No. 11
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.11.5422-5434.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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