Relative Comparison of Catalytic Characteristics of human Foamy Virus and hIV-1 Integrases

Knyazhanskaya, Ekaterina; Smolov, M. A.; Кондрашина, О. В.; Gottikh, Marina

doi:10.32607/20758251-2009-1-2-78-80

Cited by 9 publications

(8 citation statements)

References 3 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…All HIV-1 and PFV integrases carrying an N-terminal His 6 -tag were expressed and purified as previously described 51 . All Ku70 proteins carrying N-terminal GST-tag and GST alone were purified as previously described 49 .…”

Section: Methodsmentioning

confidence: 99%

Characterization of HIV-1 integrase interaction with human Ku70 protein and initial implications for drug targeting

Anisenko

Knyazhanskaya

Zalevsky

et al. 2017

Sci Rep

View full text Add to dashboard Cite

Human Ku70/Ku80 protein is known to influence HIV-1 replication. One of the possible reasons may be the protection of integrase from proteasomal degradation by Ku70 subunit. We demonstrated that recombinant HIV-1 integrase and Ku70 form a stable complex, while no interaction of Ku70 with integrase from prototype foamy virus was observed. By analyzing protein subdomains we determined two binding sites in the structure of both Ku70 and integrase: the 51–160 a.a. region of integrase interacts with residues 251–438 of Ku70, whereas Ku70 N-terminal domain (1–250 a.a.) contacts an α6-helix in the 200–220 a.a. integrase region. Single substitutions within integrase (E212A or L213A) block the interaction with Ku70 thus indicating that the binding site formed by the 200–220 a.a. integrase region is crucial for complex formation. E212A/L213A substitutions decreased the integrase capacity to bind Ku70 in HEK293T cells. A conjugate of 2′-ОMe-GGUUUUUGUGU oligonucleotide with eosin is shown by molecular modeling to shield integrase residues E212/L213 and is effective in blocking complex formation of Ku70 with integrase what makes the complex between α6-helix and Ku70(1–250) a possible target for drug development.

show abstract

Section: Methodsmentioning

confidence: 99%

Characterization of HIV-1 integrase interaction with human Ku70 protein and initial implications for drug targeting

Anisenko

Knyazhanskaya

Zalevsky

et al. 2017

Sci Rep

View full text Add to dashboard Cite

show abstract

“…A tetramer consists of asymmetrical dimers, each of those interacts with one terminus of viral DNA and performs its integration into cellular DNA [ 28 ]. The comparative study of the catalytic characteristics of the IN of the human foamy virus and HIV-1 revealed a considerable similarity in the functioning of these two enzymes [ 29 , 30 ].…”

Section: Integrase Structure and Integration Mechanismmentioning

confidence: 99%

Clinical Use of Inhibitors of HIV-1 Integration: Problems and Prospects

2011

Self Cite

View full text Add to dashboard Cite

The HIV-1 integrase enzyme is responsible for one of the key stages of retroviral replication; it acts as a catalyst for the integration of viral cDNA into the cell’s genome. Inhibitors of HIV-1 integration have been under development for over 10 years; yet, only one integration inhibitor, raltegravir, has been approved for clinical use so far. Raltegravir binds two metal ions in the enzyme’s active centre and blocks one of the integration stages: the strand transfer. Unfortunately, the clinical use of raltegravir results in the development of viral resistance among some patients. Several more HIV-1 integration inhibitors are undergoing clinical trials at the moment. However, the structure and mechanism of action of those are similar to raltegravir, which results in the emergence of cross resistance with raltegravir. The present review is focused on the history of the development and clinical trials of raltegravir and its analogues, the problems connected with the emergence of viral resistance to integration inhibitors, and the prospect of their future clinical use.

show abstract

“…Recombinant proteins expression and purification: GST-mCer-IN and His 6 -Ku70-tRFP were expressed and purified in the same way as His 6 -IN and GST-Ku70, respectively, as previously described in [ 29 , 30 ]. GST-mCer was produced and purified in the same way as GST-mCer-IN.…”

Section: Methodsmentioning

confidence: 99%

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

et al. 2020

View full text Add to dashboard Cite

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.

show abstract

Relative Comparison of Catalytic Characteristics of human Foamy Virus and hIV-1 Integrases

Cited by 9 publications

References 3 publications

Characterization of HIV-1 integrase interaction with human Ku70 protein and initial implications for drug targeting

Characterization of HIV-1 integrase interaction with human Ku70 protein and initial implications for drug targeting

Clinical Use of Inhibitors of HIV-1 Integration: Problems and Prospects

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

Contact Info

Product

Resources

About