A Fluorescent Assay to Search for Inhibitors of HIV-1 Integrase Interactions with Human Ku70 Protein, and Its Application for Characterization of Oligonucleotide Inhibitors

Galkin, Simon; Rozina, Anna; Zalevsky, Arthur; Gottikh, Marina; Anisenko, Andrey

doi:10.3390/biom10091236

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A Fluorescent Assay to Search for Inhibitors of HIV-1 Integrase Interactions with Human Ku70 Protein, and Its Application for Characterization of Oligonucleotide Inhibitors

^1,†

^1,†,

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Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, 119992 Moscow, Russia

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, 117997 Moscow, Russia

Chemistry Department, Lomonosov Moscow State University, 119992 Moscow, Russia

⁴

Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119992 Moscow, Russia

Author to whom correspondence should be addressed.

^†

These authors contributed equally to this paper.

Biomolecules 2020, 10(9), 1236; https://doi.org/10.3390/biom10091236

Received: 23 July 2020 / Revised: 21 August 2020 / Accepted: 24 August 2020 / Published: 25 August 2020

(This article belongs to the Special Issue Nucleic Acids, Structure and Modeling)

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Abstract

The search for compounds that can inhibit the interaction of certain viral proteins with their cellular partners is a promising trend in the development of antiviral drugs. We have previously shown that binding of HIV-1 integrase with human Ku70 protein is essential for viral replication. Here, we present a novel, cheap, and fast assay to search for inhibitors of these proteins’ binding based on the usage of genetically encoded fluorescent tags linked to both integrase and Ku70. Using this approach, we have elucidated structure-activity relationships for a set of oligonucleotide conjugates with eosin and shown that their inhibitory activity is primarily achieved through interactions between the conjugate nucleic bases and integrase. Molecular modeling of HIV-1 integrase in complex with the conjugates suggests that they can shield E212/L213 residues in integrase, which are crucial for its efficient binding to Ku70, in a length-dependent manner. Using the developed system, we have found the 11-mer phosphorothioate bearing 3’-end eosin-Y to be the most efficient inhibitor among the tested conjugates. View Full-Text

Keywords: HIV-1 integrase; Ku70; oligonucleotide inhibitors; protein-protein interaction inhibitors; fluorescence assay; fluorescent tags; high-throughput screening

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Galkin, S.; Rozina, A.; Zalevsky, A.; Gottikh, M.; Anisenko, A. A Fluorescent Assay to Search for Inhibitors of HIV-1 Integrase Interactions with Human Ku70 Protein, and Its Application for Characterization of Oligonucleotide Inhibitors. Biomolecules 2020, 10, 1236. https://doi.org/10.3390/biom10091236

AMA Style

Galkin S, Rozina A, Zalevsky A, Gottikh M, Anisenko A. A Fluorescent Assay to Search for Inhibitors of HIV-1 Integrase Interactions with Human Ku70 Protein, and Its Application for Characterization of Oligonucleotide Inhibitors. Biomolecules. 2020; 10(9):1236. https://doi.org/10.3390/biom10091236

Chicago/Turabian Style

Galkin, Simon, Anna Rozina, Arthur Zalevsky, Marina Gottikh, and Andrey Anisenko. 2020. "A Fluorescent Assay to Search for Inhibitors of HIV-1 Integrase Interactions with Human Ku70 Protein, and Its Application for Characterization of Oligonucleotide Inhibitors" Biomolecules 10, no. 9: 1236. https://doi.org/10.3390/biom10091236

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A Fluorescent Assay to Search for Inhibitors of HIV-1 Integrase Interactions with Human Ku70 Protein, and Its Application for Characterization of Oligonucleotide Inhibitors

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