Kinetic investigation reveals an HIV-1 Nef-dependent increase in AP-2 recruitment and productivity at endocytic sites

Mol Biol Cell. 2024 Jan 1;35(1):ar9. doi: 10.1091/mbc.E23-04-0126. Epub 2023 Nov 8.

Abstract

The HIV-1 accessory protein Nef hijacks clathrin adaptors to degrade or mislocalize host proteins involved in antiviral defenses. Here, using quantitative live-cell microscopy in genome-edited Jurkat cells, we investigate the impact of Nef on clathrin-mediated endocytosis (CME), a major pathway for membrane protein internalization in mammalian cells. Nef is recruited to CME sites on the plasma membrane, and this recruitment is associated with an increase in the recruitment and lifetime of the CME coat protein AP-2 and the late-arriving CME protein dynamin2. Furthermore, we find that CME sites that recruit Nef are more likely to recruit dynamin2 and transferrin, suggesting that Nef recruitment to CME sites promotes site maturation to ensure high efficiency in host protein downregulation. Implications of these observations for HIV-1 infection are discussed.

MeSH terms

  • Animals
  • Cell Membrane / metabolism
  • Clathrin* / metabolism
  • Endocytosis* / physiology
  • HIV-1* / metabolism
  • Humans
  • Jurkat Cells
  • Membrane Proteins / metabolism
  • nef Gene Products, Human Immunodeficiency Virus* / metabolism

Substances

  • Clathrin
  • Membrane Proteins
  • nef protein, Human immunodeficiency virus 1
  • nef Gene Products, Human Immunodeficiency Virus