title: HIV-1 Capsid Stabilization Assay.
abstract: The stability of the HIV-1 core in the cytoplasm is crucial for productive HIV-1 infection. Mutations that stabilize or destabilize the core showed defects in HIV-1 reverse transcription and infection. We developed a novel and simple assay to measure stability of in vitro-assembled HIV-1 CA-NC complexes. This assay allowed us to demonstrate that cytosolic extracts strongly stabilize the HIV-1 core (Fricke et al., J Virol 87:10587-10597, 2013). By using our novel assay, one can measure the ability of different drugs to modulate the stability of in vitro-assembled HIV-1 CA-NC complexes, such as PF74, CAP-1, IXN-053, cyclosporine A, Bi2, and the peptide CAI. We also found that purified CPSF6 (1-321) protein stabilizes in vitro-assembled HIV-1 CA-NC complexes (Fricke et al., J Virol 87:10587-10597, 2013). Here we describe in detail the use of this capsid stability assay. We believe that our assay can be a powerful tool to assess HIV-1 capsid stability in vitro.
authors.affiliation: Department of Microbiology and Immunology, Albert Einstein College of Medicine, 1301 Morris Park-Price Center 501, Bronx, NY, 10461, USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Fricke
authors.affiliation: Department of Microbiology and Immunology, Albert Einstein College of Medicine, 1301 Morris Park-Price Center 501, Bronx, NY, 10461, USA. Felipe.Diaz-Griffero@einstein.yu.edu.
authors.forename: Felipe
authors.initials: F
authors.lastname: Diaz-Griffero
mesh_terms: D019380:Anti-HIV Agents / Q000494:pharmacology; D015153:Blotting, Western / Q000379:methods; D002213:Capsid / Q000737:chemistry* / Q000187:drug effects; D036022:Capsid Proteins / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D015658:HIV Infections / Q000188:drug therapy / Q000821:virology*; D015497:HIV-1 / Q000737:chemistry* / Q000187:drug effects; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods; D055550:Protein Stability / Q000187:drug effects; D014461:Ultracentrifugation / Q000379:methods; D057646:Virus Uncoating / Q000187:drug effects
chemical_list: D019380:Anti-HIV Agents; D036022:Capsid Proteins
keywords: Capsid; Core; HIV-1; Stability; Uncoating