21047958
not annotated - annotated - LINNAEUS only
Stepwise loss of fluorescent core protein V from human adenovirus during entry into cells.
Human adenoviruses (Ads) replicate and assemble particles in the nucleus. They organize a linear double-strand DNA genome into a condensed core with about 180 nucleosomes, by the viral proteins VII (pVII), pX, and pV attaching the DNA to the capsid. Using reverse genetics, we generated a novel, nonconditionally replicating Ad reporter by inserting green fluorescent protein (GFP) at the amino terminus of pV. Purified Ad2-GFP-pV virions had an oversized complete genome and incorporated about 38 GFP-pV molecules per virion, which is about 25% of the pV levels in Ad2. GFP-pV cofractionated with the DNA core, like pV, and newly synthesized GFP-pV had a subcellular localization indistinguishable from that of pV, indicating that GFP-pV is a valid reporter for pV. Ad2-GFP-pV completed the replication cycle, although at lower yields than Ad2. Incoming GFP-pV (or pV) was not imported into the nucleus. Virions lost GFP-pV at two points during the infection process: at entry into the cytosol and at the nuclear pore complex, where capsids disassemble. Disassembled capsids, positive for the conformation-specific antihexon antibody R70, were devoid of GFP-pV. The loss of GFP-pV was reduced by the macrolide antibiotic leptomycin B (LMB), which blocks nuclear export and adenovirus attachment to the nuclear pore complex. LMB inhibited the appearance of R70 epitopes on Ad2 and Ad2-GFP-pV, indicating that the loss of GFP-pV from Ad2-GFP-pV is an authentic step in the adenovirus uncoating program. Ad2-GFP-pV is genetically complete and hence enables detailed analyses of infection and spreading dynamics in cells and model organisms or assessment of oncolytic adenoviral potential.
Ann file
T1 Species 49 65 human adenovirus
T2 Species 93 111 Human adenoviruses
T3 Species 1367 1377 adenovirus
N3 Reference T3 Taxonomy:10535
T4 Species 1565 1575 adenovirus
N4 Reference T4 Taxonomy:10535
T5 Species 1758 1768 adenoviral
N5 Reference T5 Taxonomy:10535
T6 Species 113 116 Ads
N6 Reference T6 Taxonomy:10535
N2 Reference T2 Taxonomy:1907210 Human adenovirus sp.
N1 Reference T1 Taxonomy:1907210 Human adenovirus sp.
#1 AnnotatorNotes T3 TODO check
#2 AnnotatorNotes T4 TODO check
#3 AnnotatorNotes T5 TODO check