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Journal of Virology, December 2007, p. 13235-13241, Vol. 81, No. 23
0022-538X/07/$08.00+0     doi:10.1128/JVI.00204-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Identification of an APOBEC3G Binding Site in Human Immunodeficiency Virus Type 1 Vif and Inhibitors of Vif-APOBEC3G Binding{triangledown}

Andrew Mehle,1,2 Heather Wilson,1,2 Chengsheng Zhang,1,2 Andrew Jay Brazier,1,2 Mark McPike,1,2 Erez Pery,1,2 and Dana Gabuzda1,3*

Department of Cancer Immunology and AIDS, Dana Farber Cancer Institute, Boston, Massachusetts 02115,1 Departments of Pathology,2 Neurology, Harvard Medical School, Boston, Massachusetts 021153

Received 29 January 2007/ Accepted 11 September 2007

The APOBEC3 cytidine deaminases are potent antiviral factors that restrict replication of human immunodeficiency virus type 1 (HIV-1). HIV-1 Vif binds APOBEC3G and APOBEC3F and targets these proteins for ubiquitination by forming an E3 ubiquitin ligase with cullin 5 and elongins B and C. The N-terminal region of Vif is required for APOBEC3G binding, but the binding site(s) is unknown. To identify the APOBEC3G binding site in Vif, we established a scalable binding assay in a format compatible with development of high-throughput screens. In vitro binding assays using recombinant proteins identified Vif peptides and monoclonal antibodies that inhibit Vif-APOBEC3G binding and suggested involvement of Vif residues 33 to 83 in APOBEC3G binding. Cell-based binding assays confirmed these results and demonstrated that residues 40 to 71 in the N terminus of Vif contain a nonlinear binding site for APOBEC3G. Mutation of the highly conserved residues His42/43 but not other charged residues in this region inhibited Vif-APOBEC3G binding, Vif-mediated degradation of APOBEC3G, and viral infectivity. In contrast, mutation of these residues had no significant effect on Vif binding and degradation of APOBEC3F, suggesting a differential requirement for His42/43 in Vif binding to APOBEC3G and APOBEC3F. These results identify a nonlinear APOBEC3 binding site in the N terminus of Vif and demonstrate that peptides or antibodies directed against this region can inhibit Vif-APOBEC3G binding, validating the Vif-APOBEC3 interface as a potential drug target.

* Corresponding author. Mailing address: Dana-Farber Cancer Institute, JF816, 44 Binney St., Boston, MA 02115. Phone: (617) 632-2154. Fax: (617) 632-3113. E-mail: dana_gabuzda{at}dfci.harvard.edu

{triangledown} Published ahead of print on 26 September 2007.

Journal of Virology, December 2007, p. 13235-13241, Vol. 81, No. 23
0022-538X/07/$08.00+0     doi:10.1128/JVI.00204-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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