Article (Scientific journals)
Morphogenesis of a Highly Replicative EGFPVP22 Recombinant Marek’s Disease Virus in Cell Culture
Denesvre, Caroline; Blondeau, Caroline; Lemesle, Monique et al.
2007In Journal of Virology
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Keywords :
Marek's disease virus; morphogenesis; in vitro
Abstract :
[en] Marek’s disease virus (MDV) is an alphaherpesvirus for which infection is strictly cell associated in permissive cell culture systems. In contrast to most other alphaherpesviruses, no comprehensive ultrastructural study has been published to date describing the different stages of MDV morphogenesis. To circumvent problems linked to nonsynchronized infection and low infectivity titers, we generated a recombinant MDV expressing an enhanced green fluorescent protein fused to VP22, a major tegument protein that is not implicated in virion morphogenesis. Growth of this recombinant virus in cell culture was decreased threefold compared to that of the parental Bac20 virus, but this mutant was still highly replicative. The recombinant virus allowed us to select infected cells by cell-sorting cytometry at late stages of infection for subsequent transmission electron microscopy analysis. Under these conditions, all of the stages of assembly and virion morphogenesis could be observed except extracellular enveloped virions, even at the cell surface. We observed 10-fold fewer naked cytoplasmic capsids than nuclear capsids, and intracellular enveloped virions were very rare. The partial envelopment of capsids in the cytoplasm supports the hypothesis of the acquisition of the final envelope in this cellular compartment. We demonstrate for the first time that, compared to other alphaherpesviruses, MDV seems deficient in three crucial steps of viral morphogenesis, i.e., release from the nucleus, secondary envelopment, and the exocytosis process. The discrepancy between the efficiency with which this MDV mutant spreads in cell culture and the relatively inefficient process of its envelopment and virion release raises the question of the MDV cell-to-cell spreading mechanism.
Disciplines :
Life sciences: Multidisciplinary, general & others
Author, co-author :
Denesvre, Caroline;  Institut Scientifique de Recherche Agronomique - INRA > Sante Animale > Laboratoire de Virologie Moleculaire, UR1282, IASP, Nouzilly
Blondeau, Caroline ;  Institut Scientifique de Recherche Agronomique - INRA > Sante Animale > Laboratoire de Virologie Moleculaire, UR1282, IASP, Nouzilly
Lemesle, Monique;  Universite Francois Rabelais, Tours > INSERM ERI 19, Faculte de Medecine & CHRU
Le Vern, Yves;  Institut Scientifique de Recherche Agronomique - INRA > Sante Animale > Service Commun de Cytometrie, Nouzilly
Vautherot, Daniele;  Institut Scientifique de Recherche Agronomique - INRA > Sante Animale > Laboratoire de Virologie Moleculaire, UR1282, IASP, Nouzilly
Roingeard, Philippe;  Universite Francois Rabelais, Tours > INSERM ERI 19, Faculte de Medecine & CHRU
Vautherot, Jean-Francois;  Institut Scientifique de Recherche Agronomique - INRA > Sante Animale > Laboratoire de Virologie Moleculaire, UR1282, IASP, Nouzilly
Language :
English
Title :
Morphogenesis of a Highly Replicative EGFPVP22 Recombinant Marek’s Disease Virus in Cell Culture
Publication date :
2007
Journal title :
Journal of Virology
ISSN :
0022-538X
eISSN :
1098-5514
Publisher :
American Society for Microbiology (ASM), Washington, United States - District of Columbia
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 13 March 2015

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