Dynamic Oligomerization of Integrase Orchestrates HIV Nuclear Entry

Sci Rep. 2016 Nov 10;6:36485. doi: 10.1038/srep36485.

Abstract

Nuclear entry is a selective, dynamic process granting the HIV-1 pre-integration complex (PIC) access to the chromatin. Classical analysis of nuclear entry of heterogeneous viral particles only yields averaged information. We now have employed single-virus fluorescence methods to follow the fate of single viral pre-integration complexes (PICs) during infection by visualizing HIV-1 integrase (IN). Nuclear entry is associated with a reduction in the number of IN molecules in the complexes while the interaction with LEDGF/p75 enhances IN oligomerization in the nucleus. Addition of LEDGINs, small molecule inhibitors of the IN-LEDGF/p75 interaction, during virus production, prematurely stabilizes a higher-order IN multimeric state, resulting in stable IN multimers resistant to a reduction in IN content and defective for nuclear entry. This suggests that a stringent size restriction determines nuclear pore entry. Taken together, this work demonstrates the power of single-virus imaging providing crucial insights in HIV replication and enabling mechanism-of-action studies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Active Transport, Cell Nucleus / physiology
  • Cell Line
  • Cell Nucleus / metabolism
  • Chromatin / metabolism
  • Fluorescence Resonance Energy Transfer
  • HIV Integrase / chemistry
  • HIV Integrase / genetics
  • HIV Integrase / metabolism*
  • HIV-1 / enzymology
  • HIV-1 / physiology*
  • HeLa Cells
  • Humans
  • Intercellular Signaling Peptides and Proteins / metabolism
  • Microscopy, Confocal
  • Nuclear Pore / metabolism
  • Protein Multimerization

Substances

  • Chromatin
  • Intercellular Signaling Peptides and Proteins
  • lens epithelium-derived growth factor
  • HIV Integrase
  • p31 integrase protein, Human immunodeficiency virus 1