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Journal of Virology, February 2002, p. 1959-1970, Vol. 76, No. 4
0022-538X/01/$04.00+0     DOI: 10.1128/JVI.76.4.1959-1970.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Characterization of Marek's Disease Virus Serotype 1 (MDV-1) Deletion Mutants That Lack UL46 to UL49 Genes: MDV-1 UL49, Encoding VP22, Is Indispensable for Virus Growth

Fabien Dorange,1 B. Karsten Tischer,2 Jean-François Vautherot,1* and Nikolaus Osterrieder2

Laboratoire de Virologie Moléculaire, Station de Pathologie Aviaire et de Parasitologie, Institut National de la Recherche Agronomique de Tours, 37380 Nouzilly, France,1 Institute of Molecular Biology, Friedrich-Loeffler-Institutes, Federal Research Center for Virus Diseases of Animals, D-17498 Insel Riems, Germany2

Received 9 July 2001/ Accepted 1 November 2001

Experiments were conducted to investigate the roles of Marek's disease virus serotype 1 (MDV-1) major tegument proteins VP11/12, VP13/14, VP16, and VP22 in viral growth in cultured cells. Based on a bacterial artificial chromosome clone of MDV-1 (BAC20), mutant viruses were constructed in which the MDV-1 homologs of UL46, UL47, UL48, or UL49 were deleted alone and in various combinations. It could be demonstrated that the UL46, UL47, and UL48 genes are dispensable for MDV-1 growth in chicken embryonic skin and quail muscle QM7 cells, although the generated virus mutants exhibited reduced plaque sizes in all cell types investigated. In contrast, a UL49-negative MDV-1 (20{Delta}49) and a UL48-UL49 (20{Delta}48-49) doubly negative mutant were not able to produce MDV-1-specific plaques on either cell type. It was confirmed that this growth restriction is dependent on the absence of VP22 expression, because growth of these mutant viruses could be partially restored on cells that were cotransfected with a UL49 expression plasmid. In addition, we were able to demonstrate that cell-to-cell spread of MDV-1 conferred by VP22 is dependent on the expression of amino acids 37 to 187 of MDV-1 VP22, because expression plasmids containing MDV-1 UL49 mutant genes with deletions of amino acids 1 to 37 or 188 to 250 were still able to restore partial growth of the 20{Delta}49 and 20{Delta}48-49 viruses. These results demonstrate for the first time that an alphaherpesvirus UL49-homologous gene is essential for virus growth in cell culture.

* Corresponding author. Mailing address: Laboratoire de Virologie Moléculaire, Station de Pathologie Aviaire et de Parasitologie, Centre INRA de Tours, 37380 Nouzilly, France. Phone: 33-247427986. Fax: 33-247427774. E-mail: jfvauthe{at}tours.inra.fr.

Journal of Virology, February 2002, p. 1959-1970, Vol. 76, No. 4
0022-538X/01/$04.00+0     DOI: 10.1128/JVI.76.4.1959-1970.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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