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Journal of Virology, December 2006, p. 11710-11722, Vol. 80, No. 23
0022-538X/06/$08.00+0     doi:10.1128/JVI.01038-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Inhibition of Formula-Primed Reverse Transcription by Human APOBEC3G during Human Immunodeficiency Virus Type 1 Replication{triangledown}

Fei Guo,1 Shan Cen,1,2 Meijuan Niu,1 Jenan Saadatmand,1,2 and Lawrence Kleiman1,2,3*

Lady Davis Institute for Medical Research and McGill AIDS Centre, Jewish General Hospital,1 Departments of Medicine,2 Microbiology & Immunology, McGill University, Montreal, Quebec, Canada H3T 1E23

Received 20 June 2006/ Accepted 15 September 2006

Cells are categorized as being permissive or nonpermissive according to their ability to produce infectious human immunodeficiency virus type 1 (HIV-1) lacking the viral protein Vif. Nonpermissive cells express the human cytidine deaminase APOBEC3G (hA3G), and Vif has been shown to bind to APOBEC3G and facilitate its degradation. Vif-negative HIV-1 virions produced in nonpermissive cells incorporate hA3G and have a severely reduced ability to produce viral DNA in newly infected cells. While it has been proposed that the reduction in DNA production is due to hA3G-facilitated deamination of cytidine, followed by DNA degradation, we provide evidence here that a decrease in the synthesis of the DNA by reverse transcriptase may account for a significant part of this reduction. During the infection of cells with Vif-negative HIV-1 produced from 293T cells transiently expressing hA3G, much of the inhibition of early (≥50% reduction) and late (≥95% reduction) viral DNA production, and of viral infectivity (≥95% reduction), can occur independently of DNA deamination. The inhibition of the production of early minus-sense strong stop DNA is also correlated with a similar inability of tRNA3Lys to prime reverse transcription. A similar reduction in tRNA3Lys priming and viral infectivity is also seen in the naturally nonpermissive cell H9, albeit at significantly lower levels of hA3G expression.

* Corresponding author. Mailing address: Lady Davis Institute for Medical Research, Jewish General Hospital, 3755 Cote St. Catherine Rd., Montreal, Quebec, Canada H3T 1E2. Phone: (514) 340-8260. Fax: (514) 340-7502. E-mail: lawrence.kleiman{at}mcgill.ca.

{triangledown} Published ahead of print on 13 September 2006.

Journal of Virology, December 2006, p. 11710-11722, Vol. 80, No. 23
0022-538X/06/$08.00+0     doi:10.1128/JVI.01038-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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