Characterization of herpes simplex virus-containing organelles by subcellular fractionation: Role for organelle acidification in assembly of infectious particles

Citation
Ca. Harley et al., Characterization of herpes simplex virus-containing organelles by subcellular fractionation: Role for organelle acidification in assembly of infectious particles, J VIROLOGY, 75(3), 2001, pp. 1236-1251
Citations number
83
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Microbiology
Journal title
JOURNAL OF VIROLOGY
ISSN journal
0022-538X → ACNP
Volume
75
Issue
3
Year of publication
2001
Pages
1236 - 1251
Database
ISI
SICI code
0022-538X(200102)75:3<1236:COHSVO>2.0.ZU;2-U
Abstract
The cytoplasmic compartments occupied by exocytosing herpes simplex virus ( HSV) are poorly defined. It is unclear which organelles contain the majorit y of trafficking,virions and which are occupied by virions on a productive rather than defective assembly pathway. These problems are compounded by th e bet that HSV-infected cells produce virus continuously over many hours. A ll stages in viral assembly and export therefore coexist, making it impossi ble to determine the sequence of events and their kinetics. To address thes e problems, we have established assays to monitor the presence of capsids a nd em eloped virions in cell extracts and prepared HSV-containing organelle s from normally infected cells and from cells undergoing a single synchroni zed wave of viral egress. We find that, in both cases, HSV particles exit t he nucleus and accumulate in organelles which cofractionate with the trans- Golgi network (TGN) and endosomes. In addition to carrying enveloped infect ious virions in their lumen, HSV-bearing organelles also displayed nonenvel oped capsids attached to their cytoplasmic surface. Neutralization of organ ellar pH by chloroquine or bafilomycin A resulted in the accumulation of no ninfectious enveloped particles. We conclude that the organelles of the TGN /endocytic network play a hey role in the assembly and trafficking of infec tious HSV.