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Journal of Virology, March 2004, p. 3170-3177, Vol. 78, No. 6
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.6.3170-3177.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Assessment of the Role of the Central DNA Flap in Human Immunodeficiency Virus Type 1 Replication by Using a Single-Cycle Replication System

Zhujun Ao, Xiaojian Yao,* and Éric A. Cohen*

Laboratoire de rétrovirologie humaine, Département de microbiologie et immunologie, Faculté de médecine, Université de Montréal, Montréal, Québec, Canada H3C 3J7

Received 17 November 2003/ Accepted 24 November 2003

In this study, reverse transcriptase (RT)- and integrase (IN)-defective human immunodeficiency virus type 1 (HIV-1) was transcomplemented with Vpr-RT-IN fusion proteins to delineate pol sequences important for HIV-1 replication. Our results reveal that a 194-bp sequence encompassing the 3'end of the IN gene and containing the central DNA flap is necessary and sufficient for efficient HIV-1 single-cycle replication in dividing and nondividing cells. Furthermore, we show that the central DNA flap enhances HIV-1 single-round replication by five- to sevenfold, primarily by facilitating nuclear import of proviral DNA. In agreement with previous reports, our data support a functional role of the central DNA flap during the early stages of HIV-1 infection.

* Corresponding author. Mailing address: University of Montreal, Microbiology and Immunology, CP 6128, Station Centre-Ville, Montreal, PQ, H3C 3J7 Canada. Phone, fax, and e-mail for Éric A. Cohen: Phone: (514) 343-5967. Fax: (514) 343-5995. E-mail: eric.cohen{at}umontreal.ca. Phone, fax, and e-mail for Xiaojian Yao: Phone: (514) 343-6280. Fax: (514) 343-5985. E-mail: xiaojian.yao{at}umontreal.ca.

Journal of Virology, March 2004, p. 3170-3177, Vol. 78, No. 6
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.6.3170-3177.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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