scholarly journals A High-throughput Cre-Lox Activated Viral Membrane Fusion Assay to Identify Inhibitors of HIV-1 Viral Membrane Fusion

10.3791/58074 ◽  
2018 ◽  
Author(s):  
Anthony M. Esposito ◽  
Alexandra Y. Soare ◽  
Foramben Patel ◽  
Namita Satija ◽  
Benjamin K. Chen ◽  
...  
Keyword(s):  
Membrane Fusion ◽  
High Throughput ◽  
Viral Membrane ◽  
Viral Membrane Fusion ◽  
Hiv 1

scholarly journals P2X1 Selective Antagonists Block HIV-1 Infection through Inhibition of Envelope Conformation-Dependent Fusion

2019 ◽  
Vol 94 (6) ◽  
Author(s):  
Alexandra Y. Soare ◽  
Hagerah S. Malik ◽  
Natasha D. Durham ◽  
Tracey L. Freeman ◽  
Raymond Alvarez ◽  
...  
Keyword(s):  
Membrane Fusion ◽  
Chronic Inflammation ◽  
Neutralizing Antibody ◽  
Extracellular Nucleotides ◽  
Productive Infection ◽  
Variable Regions ◽  
Viral Membrane ◽  
P2x1 Receptor ◽  
Viral Membrane Fusion ◽  
Hiv 1

ABSTRACT Purinergic receptors are well-established modulators of inflammatory processes, primarily through detection of extracellular nucleotides that are released by dying or infected cells. Emerging literature has demonstrated that inhibition of these inflammatory receptors can block HIV-1 productive infection and HIV-1-associated inflammation. The specificity of receptor type and mechanism of interaction has not yet been determined. Here, we characterize the inhibitory activity of P2X1 receptor antagonists, NF279 and NF449, in cell lines, primary cells, and a variety of HIV-1 envelope (Env) clades. NF279 and NF449 blocked productive infection at the level of viral membrane fusion, with a range of inhibitory activities against different HIV-1 Env isolates. A mutant virus carrying a truncation deletion of the C-terminal tail of HIV-1 Env glycoprotein 41 (gp41) showed reduced sensitivity to P2X1 antagonists, indicating that the sensitivity of inhibition by these molecules may be modulated by Env conformation. In contrast, a P2X7 antagonist, A438079, had a limited effect on productive infection and fusion. NF279 and NF449 interfered with the ability of the gp120 variable regions 1 and 2 (V1V2)-targeted broadly neutralizing antibody PG9 to block productive infection, suggesting that these drugs may antagonize HIV-1 Env at gp120 V1V2 to block viral membrane fusion. Our observations indicate that P2X1 antagonism can inhibit HIV-1 replication at the level of viral membrane fusion through interaction with Env. Future studies will probe the nature of these compounds in inhibiting HIV-1 fusion and the development of small molecules to block HIV-1 entry via this mechanism. IMPORTANCE While effective treatment can lower the severe morbidity and mortality associated with HIV-1 infection, patients infected with HIV-1 suffer from significantly higher rates of noncommunicable comorbidities associated with chronic inflammation. Emerging literature suggests a key role for P2X1 receptors in mediating this chronic inflammation, but the mechanism is still unknown. Here, we demonstrate that HIV-1 infection is reduced by P2X1 receptor antagonism. This inhibition is mediated by interference with HIV-1 Env and can impact a variety of viral clades. These observations highlight the importance of P2X1 antagonists as potential novel therapeutics that could serve to block a variety of different viral clades with additional benefits for their anti-inflammatory properties.

scholarly journals A high throughput Cre–lox activated viral membrane fusion assay identifies pharmacological inhibitors of HIV entry

Virology ◽  
2016 ◽  
Vol 490 ◽  
pp. 6-16 ◽  
Author(s):  
Anthony M. Esposito ◽  
Pamela Cheung ◽  
Talia H. Swartz ◽  
Hongru Li ◽  
Tshidi Tsibane ◽  
...  
Keyword(s):  
Membrane Fusion ◽  
High Throughput ◽  
Viral Membrane ◽  
Hiv Entry ◽  
Viral Membrane Fusion

Membrane-on-a-Chip: Microstructured Silicon/Silicon-Dioxide Chips for High-Throughput Screening of Membrane Transport and Viral Membrane Fusion

ACS Nano ◽  
2014 ◽  
Vol 8 (4) ◽  
pp. 3380-3392 ◽  
Author(s):  
Ilja Kusters ◽  
Antoine M. van Oijen ◽  
Arnold J. M. Driessen
Keyword(s):  
Silicon Dioxide ◽  
Membrane Fusion ◽  
Membrane Transport ◽  
High Throughput ◽  
High Throughput Screening ◽  
Viral Membrane ◽  
Viral Membrane Fusion ◽  
Silicon Silicon

scholarly journals Cell-to-Cell Transfer of HIV-1 via Virological Synapses Leads to Endosomal Virion Maturation that Activates Viral Membrane Fusion

2011 ◽  
Vol 10 (6) ◽  
pp. 551-562 ◽  
Author(s):  
Benjamin M. Dale ◽  
Gregory P. McNerney ◽  
Deanna L. Thompson ◽  
Wolfgang Hubner ◽  
Kevin de los Reyes ◽  
...  
Keyword(s):  
Membrane Fusion ◽  
Cell Transfer ◽  
Viral Membrane ◽  
Virological Synapses ◽  
Cell To Cell Transfer ◽  
Viral Membrane Fusion ◽  
Hiv 1

scholarly journals Purinergic Receptors: Elucidating the Role of These Immune Mediators in HIV-1 Fusion

2020 ◽  
Author(s):  
Tracey L. Freeman ◽  
Talia H. Swartz
Keyword(s):  
Membrane Fusion ◽  
Purinergic Receptors ◽  
Purinergic Receptor ◽  
Extracellular Nucleotides ◽  
Viral Membrane ◽  
Immune Mediators ◽  
Receptor Interactions ◽  
Viral Membrane Fusion ◽  
Hiv 1

Purinergic receptors are inflammatory mediators activated by extracellular nucleotides released by dying or injured cells. Several studies have described an important role for these receptors in HIV-1 entry, particularly regarding their activity on HIV-1 viral membrane fusion. Several reports identify purinergic receptor antagonists that inhibit HIV-1 membrane fusion; these drugs are suspected to act through antagonizing Env-chemokine receptor interactions. They also appear to abrogate activity of downstream mediators that potentiate activation of the NLRP3 inflammasome pathway. Here we review the literature on purinergic receptors, the drugs that inhibit their function, and the evidence implicating these receptors in HIV-1 entry.

scholarly journals Purinergic Receptors: Elucidating the Role of these Immune Mediators in HIV-1 Fusion

Viruses ◽  
2020 ◽  
Vol 12 (3) ◽  
pp. 290
Author(s):  
Tracey L. Freeman ◽  
Talia H. Swartz
Keyword(s):  
Membrane Fusion ◽  
Purinergic Receptors ◽  
Purinergic Receptor ◽  
Extracellular Nucleotides ◽  
Viral Membrane ◽  
Immune Mediators ◽  
Receptor Interactions ◽  
Viral Membrane Fusion ◽  
Hiv 1

Purinergic receptors are inflammatory mediators activated by extracellular nucleotides released by dying or injured cells. Several studies have described an important role for these receptors in HIV-1 entry, particularly regarding their activity on HIV-1 viral membrane fusion. Several reports identify purinergic receptor antagonists that inhibit HIV-1 membrane fusion; these drugs are suspected to act through antagonizing Env-chemokine receptor interactions. They also appear to abrogate activity of downstream mediators that potentiate activation of the NLRP3 inflammasome pathway. Here we review the literature on purinergic receptors, the drugs that inhibit their function, and the evidence implicating these receptors in HIV-1 entry.

scholarly journals P2X1 selective antagonists block HIV-1 infection through inhibition of envelope conformation-dependent fusion

2019 ◽  
Author(s):  
Alexandra Y. Soare ◽  
Hagerah S. Malik ◽  
Natasha D. Durham ◽  
Tracey L. Freeman ◽  
Raymond Alvarez ◽  
...  
Keyword(s):  
Membrane Fusion ◽  
Chronic Inflammation ◽  
Neutralizing Antibody ◽  
Productive Infection ◽  
Viral Membrane ◽  
P2x1 Receptor ◽  
Fusion Inhibition ◽  
Inflammatory Processes ◽  
Viral Membrane Fusion ◽  
Hiv 1

AbstractPurinergic receptors detect extracellular ATP and promote inflammatory processes. Emerging literature has demonstrated that inhibition of these proinflammatory receptors can block HIV-1 productive infection. The specificity of receptor type and mechanism of interaction has not yet been determined. Here we characterize the inhibitory activity of P2X1 receptor antagonists, NF279 and NF449 in cell lines, primary cells, and in a variety of envelope clades. NF279 and NF449 blocked productive infection at the level of viral membrane fusion with a range of inhibitory activities against different HIV-1 envelopes. A mutant virus carrying a truncation deletion of the C-terminal tail of HIV-1 envelope (Env) glycoprotein 41 (gp41) showed reduced sensitivity to P2X1 antagonists, indicating that the sensitivity of inhibition by these molecules is modulated by Env conformation. By contrast, a P2X7 antagonist, A438079, had limited effect on productive infection and fusion. Inhibition with NF449 interfered with the ability of the V1V2 targeted broadly neutralizing antibody PG9 to block productive infection, suggesting that these drugs may antagonize HIV-1 Env at gp120 V1V2 to block viral membrane fusion. Our observations indicate that P2X1 antagonism can inhibit HIV-1 replication at the level of viral membrane fusion through interaction with Env. Future studies will probe the nature of these compounds in inhibiting HIV-1 fusion and in development of a different class of small molecules to block HIV-1 entry.IMPORTANCEWhile effective treatment can lower the severe morbidity and mortality associated with HIV-1 infection, patients infected with HIV-1 suffer from significantly higher rates of non-communicable comorbidities associated with chronic inflammation. Emerging literature suggests a key role for P2X1 receptors in mediating this chronic inflammation but the mechanism is still unknown. Here, we demonstrate that HIV-1 infection is reduced by P2X1 receptor antagonism. This inhibition is mediated by interference with HIV-1 Env and can impact a variety of viral clades. These observations highlight the importance of P2X1 antagonists as potential novel therapeutics that could serve to block a variety of different viral clades with additional benefits for their anti-inflammatory properties.

scholarly journals Kinetically coupled folding of a single HIV-1 glycoprotein 41 complex in viral membrane fusion and inhibition

2015 ◽  
Vol 112 (22) ◽  
pp. E2855-E2864 ◽  
Author(s):  
Junyi Jiao ◽  
Aleksander A. Rebane ◽  
Lu Ma ◽  
Ying Gao ◽  
Yongli Zhang
Keyword(s):  
Membrane Fusion ◽  
Optical Tweezers ◽  
Host Cells ◽  
Dependent Manner ◽  
Folding Energy ◽  
Viral Membrane ◽  
Folding Intermediates ◽  
The Stability ◽  
Viral Membrane Fusion ◽  
Hiv 1

HIV-1 glycoprotein 41 (gp41) mediates viral entry into host cells by coupling its folding energy to membrane fusion. Gp41 folding is blocked by fusion inhibitors, including the commercial drug T20, to treat HIV/AIDS. However, gp41 folding intermediates, energy, and kinetics are poorly understood. Here, we identified the folding intermediates of a single gp41 trimer-of-hairpins and measured their associated energy and kinetics using high-resolution optical tweezers. We found that folding of gp41 hairpins was energetically independent but kinetically coupled: Each hairpin contributed a folding energy of ∼−23 kBT, but folding of one hairpin successively accelerated the folding rate of the next one by ∼20-fold. Membrane-mimicking micelles slowed down gp41 folding and reduced the stability of the six-helix bundle. However, the stability was restored by cooperative folding of the membrane-proximal external region. Surprisingly, T20 strongly inhibited gp41 folding by actively displacing the C-terminal hairpin strand in a force-dependent manner. The inhibition was abolished by a T20-resistant gp41 mutation. The energetics and kinetics of gp41 folding established by us provides a basis to understand viral membrane fusion, infection, and therapeutic intervention.

scholarly journals Viral membrane fusion

Virology ◽  
2015 ◽  
Vol 479-480 ◽  
pp. 498-507 ◽  
Author(s):  
Stephen C. Harrison
Keyword(s):  
Membrane Fusion ◽  
Viral Membrane ◽  
Viral Membrane Fusion
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