Reconstitution of herpes simplex virus microtubule-dependent trafficking in vitro

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

Microtubule-mediated anterograde transport of herpes simplex virus (HSV) from the neuronal cell body to the axon terminal is crucial for the spread and transmission of the virus. It is therefore of central importance to identify the cellular and viral factors responsible for this trafficking event. In previous studies, we isolated HSV-containing cytoplasmic organelles from infected cells and showed that they represent the first and only destination for HSV capsids after they emerge from the nucleus. In the present study, we tested whether these cytoplasmic compartments were capable of microtubule-dependent traffic. Organelles containing green fluorescent protein-labeled HSV capsids were isolated and found to be able to bind rhodamine-labeled microtubules polymerized in vitro. Following the addition of ATP, the HSV-associated organelles trafficked along the microtubules, as visualized by time lapse microscopy in an imaging microchamber. The velocity and processivity of trafficking resembled those seen for neurotropic herpesvirus traffic in living axons. The use of motor-specific inhibitors indicated that traffic was predominantly kinesin mediated, consistent with the reconstitution of anterograde traffic. Immunocytochemical studies revealed that the majority of HSV-containing organelles attached to the microtubules contained the trans-Golgi network marker TGN46. This simple, minimal reconstitution of microtubule-mediated anterograde trafic should facilitate and complement molecular analysis of HSV egress in vivo.

Original languageEnglish (US)
Pages (from-to)4264-4275
Number of pages12
JournalJournal of Virology
Volume80
Issue number9
DOIs
StatePublished - May 2006

Fingerprint

herpes simplex
Simplexvirus
Microtubules
microtubules
viruses
traffic
Organelles
organelles
capsid
Capsid
axons
Virus Release
trans-Golgi Network
Kinesin
kinesin
Rhodamines
Herpesviridae
virus transmission
Presynaptic Terminals
Green Fluorescent Proteins

ASJC Scopus subject areas

  • Immunology

Cite this

Reconstitution of herpes simplex virus microtubule-dependent trafficking in vitro. / Lee, Grace E.; Murray, John W.; Wolkoff, Allan W.; Wilson, Duncan W.

In: Journal of Virology, Vol. 80, No. 9, 05.2006, p. 4264-4275.

Research output: Contribution to journalArticle

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