Department of Microbiology and Physiological Systems
Cell Biology | Cellular and Molecular Physiology | Genetics | Microbial Physiology | Molecular Biology | Virology | Virus Diseases
Direct visualization of HIV-1 replication would improve our understanding of the viral life cycle. We adapted established technology and reagents to develop an imaging approach, ViewHIV, which allows evaluation of early HIV-1 replication intermediates, from reverse transcription to integration. These methods permit the simultaneous evaluation of both the capsid protein (CA) and viral DNA genome (vDNA) components of HIV-1 in both the cytosol and nuclei of single cells. ViewHIV is relatively rapid, uses readily available reagents in combination with standard confocal microscopy, and can be done with virtually any HIV-1 strain and permissive cell lines or primary cells. Using ViewHIV, we find that CA enters the nucleus and associates with vDNA in both transformed and primary cells. We also find that CA's interaction with the host polyadenylation factor, CPSF6, enhances nuclear entry and potentiates HIV-1's depth of nuclear invasion, potentially aiding the virus's integration into gene-dense regions.
Chin, Christopher R.; Perreira, Jill; Savidis, George; Portmann, Jocelyn M.; Aker, Aaron M.; Feeley, Eric M.; Smith, Miles C.; and Brass, Abraham L., "Direct Visualization of HIV-1 Replication Intermediates Shows that Capsid and CPSF6 Modulate HIV-1 Intra-nuclear Invasion and Integration" (2015). Microbiology and Physiological Systems Publications and Presentations. 14.
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