The HIV capsid mimics karyopherin engagement of FG-nucleoporins

HIV can infect non-dividing cells because the viral capsid can overcome the selective barrier of the nuclear pore complex and deliver the genome directly into the nucleus. Remarkably, the intact HIV capsid is over one thousand times greater than the size-limit prescribed by the nuclear pore’s diffusion barrier. This barrier is a phase-separated condensate in the central channel of the nuclear pore and is comprised of intrinsically-disordered nucleoporin domains enriched in phenylalanine-glycine (FG) dipeptides. Through multivalent FG-interactions, cellular karyopherins and their bound cargoes solubilise in this phase to drive nucleocytoplasmic transport. By performing an in vitro dissection of the nuclear pore complex, we show that a pocket on the surface of the HIV capsid similarly interacts with FG-motifs from multiple nucleoporins and that this interaction licenses capsids to penetrate nucleoporin condensates. This karyopherin mimicry model resolves a key conceptual challenge for the..., Fluorescence Fluctuation Spectroscopy Data:
Data were collected as described here: https://dx.doi.org/10.1021/acs.analchem.0c04250?ref=pdf. Fluorescence traces were analyzed using custom software TRISTAN (Two Reagents Incident Spectroscopic Analysis, freely available on https://github.com/lilbutsa/Tristan)
Confocal Microscopy Imaging:   
Imaging was performed with Zeiss LSM 880 inverted laser scanning confocal microscope using a 63x oil immersion objective (NA=1.4) (Leica, Bensheim, Germany). The substrate-Nup98 reaction mixes were transferred into a 12 well silicone chamber (Ibidi) on 170 ± 5 µm cover slide. Z-stacks were taken around the centres of the phase separated Nup98 condensates (position with highest diameter) with sequential imaging at 488 and 568. Images were processed using ImageJ and Matlab.
Cryo-Electron Tomography
The grids were imaged on a Talos Arctica electron microscope (Thermo Fisher Scientific) operated at 200 kV acceleration voltage. Cryo-ET data were collected wi..., Fluorescence Fluctuation Spectroscopy Data:
Fluorescence traces can be opened with TRISTAN (https://github.com/lilbutsa/Tristan)
Confocal Microscopy Imaging:
.czi files can be opened in ImageJ.
Cryo-Electron Tomography:
.mrc files can be opened in IMOD.
Other:
.pzfx files can be opened in GraphPad Prism, ## Fluorescence fluctuation spectroscopy, confocal microscopy, and cryo-elecrtron tomography data from 'The HIV capsid mimics karyopherin engagement of FG-nucleoporins'

## Description of the data and file structure

Data archive for 'Data from: The HIV capsid mimics karyopherin engagement of FG-nucleoporins'

by Dickson, C.F.*, Hertel, S.*, Ruan, J., Ariotti, N., Tuckwell, A., Li, N., Al-Izzi, S.C., Sierecki, E., Gambin, Y., Morris, R.G., Towers, G.J., Böcking, T., Jacques, D.A.

This dataset contains data and analysis files that should allow the replication of outputs demonstrating that the HIV capsid specifically engages with FG-nucleoporins, and that this interaction enable the virus to penetrate the diffusion barrier of the nuclear pore complex. These results show that the HIV capsid mimics the endogenous nuclear transport receptors (the karyopherin) to gain access to the nucleus, explaining how the virus can infect non-dividing cells.

Data are structured according to the figures ...