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

C. A. Harley, A. Dasgupta, Duncan W. Wilson

Research output: Contribution to journalArticle

94 Citations (Scopus)

Abstract

The cytoplasmic compartments occupied by exocytosing herpes simplex virus (HSV) are poorly defined. It is unclear which organelles contain the majority of trafficking virions and which are occupied by virions on a productive rather than defective assembly pathway. These problems are compounded by the fact that HSV-infected cells produce virus continuously over many hours. All stages in viral assembly and export therefore coexist, making it impossible to determine the sequence of events and their kinetics. To address these problems, we have established assays to monitor the presence of capsids and enveloped virions in cell extracts and prepared HSV-containing organelles from normally infected cells and from cells undergoing a single synchronized wave of viral egress. We find that, in both cases, HSV particles exit the nucleus and accumulate in organelles which cofractionate with the trans-Golgi network (TGN) and endosomes. In addition to carrying enveloped infectious virions in their lumen, HSV-bearing organelles also displayed nonenveloped capsids attached to their cytoplasmic surface. Neutralization of organellar pH by chloroquine or bafilomycin A resulted in the accumulation of nonifectious enveloped particles. We conclude that the organelles of the TGN/endocytic network play a key role in the assembly and trafficking of infectious HSV.

Original languageEnglish (US)
Pages (from-to)1236-1251
Number of pages16
JournalJournal of Virology
Volume75
Issue number3
DOIs
StatePublished - 2001

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herpes simplex
Simplexvirus
Organelles
acidification
organelles
fractionation
virion
Virion
viruses
trans-Golgi Network
capsid
Capsid
cells
Virus Assembly
chloroquine
endosomes
Endosomes
Chloroquine
Cell Extracts
neutralization

ASJC Scopus subject areas

  • Immunology

Cite this

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