Open AccessFluorescent protein tagging of adenoviral proteins pV and pIX reveals ‘late virion accumulation compartment’
Author(s) -
Søren Pfitzner,
Helga Hofmann-Sieber,
Jens B. Bosse,
Linda E. Franken,
Kay Grünewald,
Thomas Dobner
Publication year - 2020
Publication title -
plos pathogens
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.719
H-Index - 206
eISSN - 1553-7374
pISSN - 1553-7366
DOI - 10.1371/journal.ppat.1008588
Subject(s) - compartment (ship) , microbiology and biotechnology , capsid , biology , green fluorescent protein , organelle , cellular compartment , fluorescence microscope , protein subcellular localization prediction , immunoelectron microscopy , viral replication , transport protein , immunofluorescence , virus , virology , cell , gene , antibody , fluorescence , genetics , oceanography , physics , quantum mechanics , geology
The human adenovirus type 5 (HAdV5) causes disease of the upper and lower respiratory tract. The early steps of HAdV5 entry up to genome replication in the host nucleus have been extensively studied. However, late stages of infection remain poorly understood. Here, we set out to elucidate the spatiotemporal orchestration of late adenovirus nuclear remodeling in living cells. We generated virus mutants expressing fluorescently tagged protein IX (pIX) and protein V (pV), a capsid and viral genome associated protein, respectively. We found that during progeny virion production both proteins localize to a membrane-less, nuclear compartment, which is highly impermeable such that in immunofluorescence microscopy antibodies can hardly penetrate it. We termed this compartment ‘late virion accumulation compartment’ (LVAC). Correlation between light- and electron microscopy revealed that the LVAC contains paracrystalline arrays of viral capsids that arrange tightly packed within a honeycomb-like organization of viral DNA. Live-cell microscopy as well as FRAP measurements showed that the LVAC is rigid and restricts diffusion of larger molecules, indicating that capsids are trapped inside.