Antiviral Activity of Recombinant Ankyrin Targeted to the Capsid Domain of HIV-1 Gag Polyprotein

Overview

Journal Retrovirology

Publisher Biomed Central

Specialty Microbiology

Date 2012 Feb 22

PMID 22348230

Citations 20

Authors

Sawitree Nangola

Agathe Urvoas

Marie Valerio-Lepiniec

Wannisa Khamaikawin

Supachai Sakkhachornphop

Saw-See Hong

Pierre Boulanger

Philippe Minard

Chatchai Tayapiwatana

Affiliations

Soon will be listed here.

Abstract

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(GAG)1D4 (16.5 kDa) was isolated. Ank(GAG)1D4 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(GAG)1D4 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(GAG)1D4 in both N-myristoylated and non-N-myristoylated versions was significantly reduced compared to control cells. Ank(GAG)1D4 expression also reduced the production of MLV, a phylogenetically distant retrovirus. The Ank(GAG)1D4-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(GAG)1D4-CA with the Gag assembly and budding pathway.

Conclusions: The resistance of Ank(GAG)1D4-expressing cells to HIV-1 suggested that the CA-targeted ankyrin Ank(GAG)1D4 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.

Citing Articles

Dimeric Ankyrin with Inverted Module Promotes Bifunctional Property in Capturing Capsid to Impede HIV-1 Replication.

Juntit O, Sornsuwan K, Wisitponchai T, Lee V, Sakkhachornphop S, Yasamut U Int J Mol Sci. 2023; 24(6).

PMID: 36982337 PMC: 10048781. DOI: 10.3390/ijms24065266.

Validation of Promoters and Codon Optimization on CRISPR/Cas9-Engineered Jurkat Cells Stably Expressing αRep4E3 for Interfering with HIV-1 Replication.

Chupradit K, Sornsuwan K, Saiprayong K, Wattanapanitch M, Tayapiwatana C Int J Mol Sci. 2022; 23(23).

PMID: 36499376 PMC: 9738563. DOI: 10.3390/ijms232315049.

Designed Ankyrin Repeat Proteins: A New Class of Viral Entry Inhibitors.

Walser M, Mayor J, Rothenberger S Viruses. 2022; 14(10).

PMID: 36298797 PMC: 9611651. DOI: 10.3390/v14102242.

Specific Interaction of DARPin with HIV-1 CA Disturbs the Distribution of Gag, RNA Packaging, and Tetraspanin Remodelling in the Membrane.

Moonmuang S, Maniratanachote R, Chetprayoon P, Sornsuwan K, Thongkum W, Chupradit K Viruses. 2022; 14(4).

PMID: 35458554 PMC: 9025900. DOI: 10.3390/v14040824.

Targeting the Virus Capsid as a Tool to Fight RNA Viruses.

Hozakova L, Vokata B, Ruml T, Ulbrich P Viruses. 2022; 14(2).

PMID: 35215767 PMC: 8879806. DOI: 10.3390/v14020174.

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