Single hepatitis-B virus core capsid binding to individual nuclear pore complexes in Hela cells

Biophys J. 2006 Oct 15;91(8):3123-30. doi: 10.1529/biophysj.106.087650. Epub 2006 Jul 28.

Abstract

We investigate the interaction of hepatitis B virus capsids lacking a nuclear localization signal with nuclear pore complexes (NPCs) in permeabilized HeLa cells. Confocal and wide-field optical images of the nuclear envelope show well-spaced individual NPCs. Specific interactions of capsids with single NPCs are characterized by extended residence times of capsids in the focal volume which are characterized by fluorescence correlation spectroscopy. In addition, single-capsid-tracking experiments using fast wide-field fluorescence microscopy at 50 frames/s allow us to directly observe specific binding via a dual-color colocalization of capsids and NPCs. We find that binding occurs with high probability on the nuclear-pore ring moiety, at 44 +/- 9 nm radial distance from the central axis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Capsid / metabolism*
  • Computer Simulation
  • HeLa Cells
  • Hepatitis B virus / metabolism*
  • Humans
  • Microscopy, Fluorescence
  • Nuclear Localization Signals / metabolism
  • Nuclear Pore / metabolism*
  • Protein Binding
  • Spectrometry, Fluorescence
  • Viral Core Proteins / metabolism*

Substances

  • Nuclear Localization Signals
  • Viral Core Proteins