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Single Hepatitis-B Virus Core Capsid Binding to Individual Nuclear Pore Complexes in HeLa Cells

Yoriko Lill ^*, Markus A. Lill , Birthe Fahrenkrog , Kyrill Schwarz-Herion , Sara Paulillo , Ueli Aebi  and Bert Hecht ^*

^* Nano-Optics Group, National Competence Center for Research in Nanoscale Science, Institute of Physics; Biographics Laboratory 3R and Department of Molecular Pharmacy; and M. E. Müller Institute, Biozentrum, University of Basel, CH-4056 Basel, Switzerland

Correspondence: Address reprint requests to Bert Hecht, Nano-Optics Group, National Competence Center for Research in Nanoscale Science, Institute of Physics, University of Basel, Klingelbergstrasse 82, CH-4056 Basel, Switzerland. Fax: 41-61-267-3795; E-Mail: bert.hecht{at}unibas.ch.

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.