HIV-1 Uncoating and Nuclear Import Precede the Completion of Reverse Transcription in Cell Lines and in Primary Macrophages
Abstract
:1. Introduction
2. Materials and Methods
2.1. Plasmids
2.2. Cell Lines and Reagents
2.3. Pseudovirus Production and Characterization
2.4. Isolation, Differentiation and Treatment of MDMs
2.5. Single-Cycle Infection Assay
2.6. Fixed-Cell Imaging and Immunofluorescence Assay
2.7. Live-Cell Imaging of HIV-1 Uncoating, Nuclear Entry, and Infection
2.8. Image Acquisition
2.9. Single-Particle Tracking and Image Analysis
2.10. Statistical Analyses
3. Results
3.1. Productive HIV-1 Uncoating and Nuclear Import Precede the Completion of Reverse Transcription in Cell Lines
3.2. Reverse Transcription Continues Past HIV-1 Escape from CA-Targeting Inhibitor and Is Completed in the Nucleus of MDMs, Independent of SAMHD1 Depletion
3.3. SAMHD1 Depletion in MDMs Diminishes the Pool of Stable Post-Fusion Cores
3.4. Long-Lived HIV-1 Cores Retain CDR over the Course of Several Hours Irrespective of SAMHD1 Depletion
3.5. HIV-1 Nuclear Import in MDMs Progresses through a Loss of CDR at the Nuclear Envelope and Is Accelerated upon SAMHD1 Depletion
3.6. Improved HIV-1 Infection in SAMHD1-Depleted MDMs Correlates with a More Efficient Nuclear Reverse Transcription
3.7. Host Transcriptional Factors Are Enriched at the Sites of vDNA-Containing Nuclear HIV-1 Clusters
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Francis, A.C.; Marin, M.; Prellberg, M.J.; Palermino-Rowland, K.; Melikyan, G.B. HIV-1 Uncoating and Nuclear Import Precede the Completion of Reverse Transcription in Cell Lines and in Primary Macrophages. Viruses 2020, 12, 1234. https://doi.org/10.3390/v12111234
Francis AC, Marin M, Prellberg MJ, Palermino-Rowland K, Melikyan GB. HIV-1 Uncoating and Nuclear Import Precede the Completion of Reverse Transcription in Cell Lines and in Primary Macrophages. Viruses. 2020; 12(11):1234. https://doi.org/10.3390/v12111234
Chicago/Turabian StyleFrancis, Ashwanth C., Mariana Marin, Mathew J. Prellberg, Kristina Palermino-Rowland, and Gregory B. Melikyan. 2020. "HIV-1 Uncoating and Nuclear Import Precede the Completion of Reverse Transcription in Cell Lines and in Primary Macrophages" Viruses 12, no. 11: 1234. https://doi.org/10.3390/v12111234