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

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, m...

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Main Authors: Sawitree Nangola, Agathe Urvoas, Marie Valerio-Lepiniec, Wannisa Khamaikawin, Supachai Sakkhachornphop, Saw See Hong, Pierre Boulanger, Philippe Minard, Chatchai Tayapiwatana
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Published: 2018
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http://cmuir.cmu.ac.th/jspui/handle/6653943832/51741
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spelling th-cmuir.6653943832-517412018-09-04T06:12:01Z Antiviral activity of recombinant ankyrin targeted to the capsid domain of HIV-1 Gag polyprotein Sawitree Nangola Agathe Urvoas Marie Valerio-Lepiniec Wannisa Khamaikawin Supachai Sakkhachornphop Saw See Hong Pierre Boulanger Philippe Minard Chatchai Tayapiwatana Immunology and Microbiology Medicine 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. © 2012 Nangola et al; licensee BioMed Central Ltd. 2018-09-04T06:07:17Z 2018-09-04T06:07:17Z 2012-02-20 Journal 17424690 2-s2.0-84857126902 10.1186/1742-4690-9-17 https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84857126902&origin=inward http://cmuir.cmu.ac.th/jspui/handle/6653943832/51741
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
collection CMU Intellectual Repository
topic Immunology and Microbiology
Medicine
spellingShingle Immunology and Microbiology
Medicine
Sawitree Nangola
Agathe Urvoas
Marie Valerio-Lepiniec
Wannisa Khamaikawin
Supachai Sakkhachornphop
Saw See Hong
Pierre Boulanger
Philippe Minard
Chatchai Tayapiwatana
Antiviral activity of recombinant ankyrin targeted to the capsid domain of HIV-1 Gag polyprotein
description 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. © 2012 Nangola et al; licensee BioMed Central Ltd.
format Journal
author Sawitree Nangola
Agathe Urvoas
Marie Valerio-Lepiniec
Wannisa Khamaikawin
Supachai Sakkhachornphop
Saw See Hong
Pierre Boulanger
Philippe Minard
Chatchai Tayapiwatana
author_facet Sawitree Nangola
Agathe Urvoas
Marie Valerio-Lepiniec
Wannisa Khamaikawin
Supachai Sakkhachornphop
Saw See Hong
Pierre Boulanger
Philippe Minard
Chatchai Tayapiwatana
author_sort Sawitree Nangola
title Antiviral activity of recombinant ankyrin targeted to the capsid domain of HIV-1 Gag polyprotein
title_short Antiviral activity of recombinant ankyrin targeted to the capsid domain of HIV-1 Gag polyprotein
title_full Antiviral activity of recombinant ankyrin targeted to the capsid domain of HIV-1 Gag polyprotein
title_fullStr Antiviral activity of recombinant ankyrin targeted to the capsid domain of HIV-1 Gag polyprotein
title_full_unstemmed Antiviral activity of recombinant ankyrin targeted to the capsid domain of HIV-1 Gag polyprotein
title_sort antiviral activity of recombinant ankyrin targeted to the capsid domain of hiv-1 gag polyprotein
publishDate 2018
url https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84857126902&origin=inward
http://cmuir.cmu.ac.th/jspui/handle/6653943832/51741
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