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The Cyclosporin A Washout Assay to Detect HIV-1 Uncoating in Infected Cells

  • Amy E. Hulme
  • Thomas J. Hope
Part of the Methods in Molecular Biology book series (MIMB, volume 1087)

Abstract

Uncoating is an early step of HIV-1 replication in which the viral capsid disassembles by p24 capsid (p24CA) protein dissociating from the viral complex. Although uncoating is required for HIV-1 replication, many questions remain about the mechanism of this process as well as its impact on other steps in viral replication. Here we describe a recently developed assay to study the process of uncoating in HIV-1-infected cells. The CsA washout assay is a cell-based assay that utilizes the HIV-1 restriction factor TRIM-CypA to detect and inhibit infection of coated viral complexes. Owl monkey kidney (OMK) cells are infected with a GFP reporter virus and TRIM-CypA restriction is switched on at various times postinfection allowing the kinetics of uncoating to be monitored in infected cells. This assay also can be used to examine the effect of different viral or cellular factors on the process of uncoating.

Key words

HIV-1 Capsid Uncoating Retrovirus CsA washout assay TRIM-CypA 

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Copyright information

© Springer Science+Business Media, LLC 2014

Authors and Affiliations

  • Amy E. Hulme
    • 1
  • Thomas J. Hope
    • 2
  1. 1.Department of Cell and Molecular BiologyNorthwestern University Medical SchoolChicagoUSA
  2. 2.Departments of Infectious Diseases and Cell and Molecular Biology, Feinberg School of MedicineNorthwestern UniversityChicagoUSA

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