ArticleBiophysical journal2021
Single-virus content-mixing assay reveals cholesterol-enhanced influenza membrane fusion efficiency.
Article in Biophysical journal, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 36 citations in OpenAlex.
- Single-Virus Lipid-Mixing Study of Sendai Virus Provides Insight into Fusion Mechanism.ACS omega · 2025Article
- Single VLP lipid-mixing measurements confirm off-pathway state in dengue virus fusion mechanism.Biophysical journal · 2025Article
- Influenza viral infection at the plasma membrane is restricted by lipid composition.Journal of virology · 2025Article
- Unraveling dual fusion mechanisms in BmNPV GP64: critical roles of CARC motifs and signal peptide retention.Journal of virology · 2025Article
- Article
- Insights into IAV Replication and Lipid Metabolism in Suspension-Adapted MDCK-STAT1-KO Cells.Vaccines · 2025Article
- A Fluorogenic Pseudoinfection Assay to Probe Transfer and Distribution of Influenza Viral Contents to Target Vesicles.Analytical chemistry · 2024Article
- Single-Virus Fusion Measurements Reveal Multiple Mechanistically Equivalent Pathways for SARS-CoV-2 Entry.Journal of virology · 2023Article
- Influenza Virus Membrane Fusion Is Promoted by the Endosome-Resident Phospholipid Bis(monoacylglycero)phosphate.The journal of physical chemistry. B · 2022Article
- Measuring single-virus fusion kinetics using an assay for nucleic acid exposure.Biophysical journal · 2022Article
- Recent Developments in Single-Virus Fusion Assay.The Journal of membrane biology · 2022Review
- The constellation of cholesterol-dependent processes associated with SARS-CoV-2 infection.Progress in lipid research · 2022Review
- Modulating the Influenza A Virus-Target Membrane Fusion Interface With Synthetic DNA-Lipid Receptors.Langmuir : the ACS journal of surfaces and colloids · 2022Article
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Authors and funding
2 authors at 2 institutions in 1 country.
Funding
Abstract
To infect a cell, enveloped viruses must first undergo membrane fusion, which proceeds through a hemifusion intermediate, followed by the formation of a fusion pore through which the viral genome is transferred to a target cell. Single-virus fusion studies to elucidate the dynamics of content mixing typically require extensive fluorescent labeling of viral contents. The labeling process must be optimized depending on the virus identity and strain and can potentially be perturbative to viral fusion behavior. Here, we introduce a single-virus assay in which content-labeled vesicles are bound to unlabeled influenza A virus (IAV) to eliminate the problematic step of content-labeling virions. We use fluorescence microscopy to observe individual, pH-triggered content mixing and content-loss events between IAV and target vesicles of varying cholesterol compositions. We show that target membrane cholesterol increases the efficiency of IAV content mixing and decreases the fraction of content-mixing events that result in content loss. These results are consistent with previous findings that cholesterol stabilizes pore formation in IAV entry and limits leakage after pore formation. We also show that content loss due to hemagglutinin fusion peptide engagement with the target membrane is independent of composition. This approach is a promising strategy for studying the single-virus content-mixing kinetics of other enveloped viruses.
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