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Quantitative live-cell imaging of human immunodeficiency virus (HIV-1) assembly
Viola Baumgärtel1, Barbara Müller, Don C Lamb
1Department of Chemistry, Center for NanoScience (CeNS) and Center for Integrated Protein Science, Munich (CIPSM), Ludwig-Maximilians-Universität München, Butenandtstr. 5-13, D-81377 München, Germany. viola.baumgaertel@cup.uni-muenchen.de
Insights
Quantitative live-cell imaging advances visualize human immunodeficiency virus type 1 (HIV-1) dynamics. This review details fluorescence labeling and microscopy methods to study late-stage HIV-1 replication and virus-cell interactions.
Area of Science:
- Microbiology
- Cell Biology
- Biophysics
Background:
- Quantitative live-cell imaging is crucial for understanding dynamic biological processes.
- Fluorescence microscopy techniques offer unprecedented detail in biological investigations.
- Studying virus-cell interactions requires advanced imaging methods.
Purpose of the Study:
- To review fluorescence labeling strategies for human immunodeficiency virus type 1 (HIV-1) live cell imaging.
- To present fluorescence-based methods for visualizing virus-cell interactions.
- To elucidate the dynamics of late stages in the HIV-1 replication cycle using quantitative microscopy.
Main Methods:
- Fluorescence labeling of HIV-1 components.
- Quantitative live-cell imaging techniques.
- Confocal and super-resolution microscopy for high-resolution visualization.
Main Results:
- Detailed visualization of cytosolic interactions of the Gag protein with itself and viral RNA.
- Observation of Gag and RNA recruitment to the plasma membrane.
- Analysis of virion assembly and recruitment of cellular proteins during HIV-1 release.
Conclusions:
- Quantitative microscopy provides critical insights into late-stage HIV-1 replication dynamics.
- Fluorescence imaging is indispensable for understanding virus-cell interactions.
- Advanced imaging methods facilitate the study of viral assembly and egress.
Abstract:
Advances in fluorescence methodologies make it possible to investigate biological systems in unprecedented detail. Over the last few years, quantitative live-cell imaging has increasingly been used to study the dynamic interactions of viruses with cells and is expected to become even more indispensable in the future. Here, we describe different fluorescence labeling strategies that have been used to label HIV-1 for live cell imaging and the fluorescence based methods used to visualize individual aspects of virus-cell interactions. This review presents an overview of experimental methods and recent experiments that have employed quantitative microscopy in order to elucidate the dynamics of late stages in the HIV-1 replication cycle. This includes cytosolic interactions of the main structural protein, Gag, with itself and the viral RNA genome, the recruitment of Gag and RNA to the plasma membrane, virion assembly at the membrane and the recruitment of cellular proteins involved in HIV-1 release to the nascent budding site.
