Quantification of HIV-1 Gag Localization Within Virus Producer Cells

Methods Mol Biol. 2016:1354:165-74. doi: 10.1007/978-1-4939-3046-3_11.

Abstract

Trafficking of newly synthesized Gag protein to the plasma membrane is one of the important steps during HIV-1 assembly. It requires participation of both viral and cellular determinants. Several techniques have been used to measure the amount of Gag that is associated with plasma membrane. Here we describe a microscopy-based method to estimate the distribution of Gag protein within the producer cell. This method can be used in conjunction with other biochemical techniques to quantify the distribution of Gag within a virus-producing cell and its accumulation at the plasma membrane. Since this method is microscopy based, it allows one to quantitate Gag across the cytoplasm, from the nuclear periphery to plasma membrane, at the single-cell level.

Keywords: Gag trafficking; HIV-1 Gag; Quantitating Gag protein.

Publication types

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

MeSH terms

  • Cell Line
  • HIV Infections / metabolism*
  • HIV Infections / pathology
  • HIV Infections / virology*
  • HIV-1 / isolation & purification
  • HIV-1 / physiology*
  • Humans
  • Microscopy, Confocal / methods
  • Microscopy, Fluorescence / methods*
  • Optical Imaging / methods
  • Protein Transport
  • Software
  • gag Gene Products, Human Immunodeficiency Virus / analysis*
  • gag Gene Products, Human Immunodeficiency Virus / metabolism*

Substances

  • gag Gene Products, Human Immunodeficiency Virus