Antiviral activity of recombinant ankyrin targeted to the capsid domain of HIV-1 Gag polyprotein

Retrovirology

Chatchai Tayapiwatana²  Philippe Minard³  Pierre Boulanger¹  Saw-See Hong¹  Supachai Sakkhachornphop²  Wannisa Khamaikawin²  Marie Valerio-Lepiniec³  Agathe Urvoas³  Sawitree Nangola³ 

[1] INRA UMR-754, Retrovirus and Comparative Pathology, 50, avenue Tony Garnier, 69366 Lyon Cedex 07, France;Biomedical Technology Research Unit, National Center for GeneticEngineering and Biotechnology, National Science and Technology Development Agency at the Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand;Institut de Biochimie et de Biophysique Moléculaire et Cellulaire (IBBMC) UMR-8619, Université de Paris-Sud et CNRS, Orsay Cedex 91405, France

关键词: intracellular antiviral agent;    artificial ankyrin library;    ankyrins;    virus assembly inhibitor;    CA domain;    Gag polyprotein;    HIV-1 assembly;    HIV-1;   

Others  :  1209332
DOI  :  10.1186/1742-4690-9-17

 received in 2011-08-15, accepted in 2012-02-20,  发布年份 2012

【 摘 要 】

Background

Ankyrins are cellular mediators of a number of essential protein-protein interactions. Unlike intrabodies, ankyrins are composed of highly structured repeat modules characterized by disulfide bridge-independent folding. Artificial ankyrin molecules, designed to target viral components, might act as intracellular antiviral agents and contribute to the cellular immunity against viral pathogens such as HIV-1.

Results

A phage-displayed library of artificial ankyrins was constructed, and screened on a polyprotein made of the fused matrix and capsid domains (MA-CA) of the HIV-1 Gag precursor. An ankyrin with three modules named Ank^GAG1D4 (16.5 kDa) was isolated. Ank^GAG1D4 and MA-CA formed a protein complex with a stoichiometry of 1:1 and a dissociation constant of K_d ~ 1 μM, and the Ank^GAG1D4 binding site was mapped to the N-terminal domain of the CA, within residues 1-110. HIV-1 production in SupT1 cells stably expressing Ank^GAG1D4 in both N-myristoylated and non-N-myristoylated versions was significantly reduced compared to control cells. Ank^GAG1D4 expression also reduced the production of MLV, a phylogenetically distant retrovirus. The Ank^GAG1D4-mediated antiviral effect on HIV-1 was found to occur at post-integration steps, but did not involve the Gag precursor processing or cellular trafficking. Our data suggested that the lower HIV-1 progeny yields resulted from the negative interference of Ank^GAG1D4-CA with the Gag assembly and budding pathway.

Conclusions

The resistance of Ank^GAG1D4-expressing cells to HIV-1 suggested that the CA-targeted ankyrin Ank^GAG1D4 could serve as a protein platform for the design of a novel class of intracellular inhibitors of HIV-1 assembly based on ankyrin-repeat modules.

【 授权许可】

   
2012 Nangola et al; licensee BioMed Central Ltd.

【 图 表 】

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