Biochemical detection of capsid in the nucleus during HIV-1 infection

STAR Protoc. 2021 Feb 3;2(1):100323. doi: 10.1016/j.xpro.2021.100323. eCollection 2021 Mar 19.

Abstract

To understand the role of the HIV-1 capsid in viral replication, we developed a protocol to biochemically track capsid in the nucleus during infection. To this end, we separated HIV-1-infected cells into nuclear and cytosolic fractions. Fractions were analyzed by western blotting for HIV-1 capsid content as well as for nuclear and cytosolic markers to assess the bona fide origin of the fractions. This protocol can be applied in both cycling and non-cycling human cells. For complete details on the use and execution of this protocol, please refer to Selyutina et al. (2020a).

Keywords: Cell biology; Molecular biology; Protein biochemistry.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • A549 Cells
  • Animals
  • Capsid / metabolism*
  • Cell Nucleus* / metabolism
  • Cell Nucleus* / pathology
  • Cell Nucleus* / virology
  • Cytosol* / metabolism
  • Cytosol* / pathology
  • Cytosol* / virology
  • Dogs
  • HEK293 Cells
  • HIV Infections* / metabolism
  • HIV Infections* / pathology
  • HIV-1 / physiology*
  • HeLa Cells
  • Humans
  • Virus Replication*