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Journal of Virology, February 2005, p. 1389-1396, Vol. 79, No. 3
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.3.1389-1396.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

The Ataxia Telangiectasia-Mutated and Rad3-Related Protein Is Dispensable for Retroviral Integration

Jason L. DeHart,1 Joshua L. Andersen,1 Erik S. Zimmerman,1 Orly Ardon,1 Dong Sung An,2 Jana Blackett,1 Baek Kim,3 and Vicente Planelles1*

Division of Cellular Biology and Immunology, Department of Pathology, University of Utah School of Medicine, Salt Lake City, Utah,1 Division of Hematology-Oncology, Department of Medicine, University of California- Los Angeles School of Medicine, Los Angeles, California; and Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York,2 Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York3

Received 9 June 2004/ Accepted 10 September 2004

Integration into the host cell DNA is an essential part of the retroviral life cycle and is required for the productive replication of a retrovirus. Retroviral integration involves cleavage of the host DNA and insertion of the viral DNA, forming an integration intermediate that contains two gaps, each with a viral 5' flap. The flaps are then removed, and the gap is filled by as yet unidentified nuclease and polymerase activities. It is thought that repair of these gaps flanking the site of retroviral integration is achieved by host DNA repair machinery. The ATM and Rad3-related protein (ATR) is a member of the phosphatidylinositol 3 kinase-related family of protein kinases that play a major role in sensing and triggering repair of DNA lesions in mammalian cells. In an effort to examine the role of ATR in retroviral integration, we used RNA interference to selectively downregulate ATR and measured integration efficiency. In addition, we examined the possible role that Vpr may play in enhancing integration and, in particular, whether activation of ATR by Vpr (Roshal et al., J. Biol. Chem. 278:25879-25886, 2003) will favor human immunodeficiency virus type 1 integration. We conclude that cells in which ATR has been depleted are competent for retroviral integration. We also conclude that the presence of Vpr as a virion-bound protein does not enhance integration of a lentivirus vector in dividing cells.

* Corresponding author. Mailing address: Department of Pathology, University of Utah School of Medicine, 30 N 1900 East, SOM 5C210, Salt Lake City, UT 84132. Phone: (801) 581-8655. Fax: (801) 581-8655. E-mail: vicente.planelles{at}path.utah.edu.

Journal of Virology, February 2005, p. 1389-1396, Vol. 79, No. 3
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.3.1389-1396.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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