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Journal of Virology, December 2001, p. 11307-11318, Vol. 75, No. 23
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.23.11307-11318.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Glycoproteins E and I of Marek's Disease Virus
Serotype 1 Are Essential for Virus Growth in Cultured Cells
Daniel
Schumacher,1
B. Karsten
Tischer,1
Sanjay M.
Reddy,2 and
Nikolaus
Osterrieder1,*
Institute of Molecular Biology,
Friedrich-Loeffler-Institutes, Federal Research Centre for Virus
Diseases of Animals, D-17498 Insel Riems,
Germany,1 and Avian Disease and Oncology
Laboratory, United States Department of Agriculture, East Lansing,
Michigan 488232
Received 18 June 2001/Accepted 10 August 2001
The role of glycoprotein E (gE) and gI of Marek's disease virus
serotype 1 (MDV-1) for growth in cultured cells was investigated. MDV-1
mutants lacking either gE (20
gE), gI (20
gI), or both gE and gI
(20
gEI) were constructed by recE/T-mediated mutagenesis of a
recently established infectious bacterial artificial chromosome (BAC)
clone of MDV-1 (D. Schumacher, B. K. Tischer, W. Fuchs, and N. Osterrieder, J. Virol. 74:11088-11098, 2000).
Deletion of either gE or gI, which form a complex in MDV-1-infected
cells, resulted in the production of virus progeny that were unable to spread from cell to cell in either chicken embryo fibroblasts or quail
muscle cells. This was reflected by the absence of virus plaques and
the detection of only single infected cells after transfection, even
after coseeding of transfected cells with uninfected cells. In
contrast, growth of rescuant viruses, in which the deleted glycoprotein
genes were reinserted by homologous recombination, was
indistinguishable from that of parental BAC20 virus. In addition, the
20
gE mutant virus was able to spread from cell to cell when cotransfected into chicken embryo fibroblasts with an expression plasmid encoding MDV-1 gE, and the 20
gI mutant virus exhibited cell-to-cell spread capability after cotransfection with a gI expression plasmid. The 20
gEI mutant virus, however, was not able to
spread in the presence of either a gE or gI expression plasmid, and
only single infected cells were detected by indirect immunofluorescence. The results reported here demonstrate for the first
time that both gE and gI are absolutely essential for cell-to-cell
spread of a member of the Alphaherpesvirinae.
*
Corresponding author. Mailing address: Institute of
Molecular Biology, Friedrich-Loeffler-Institutes, Federal Research
Centre for Virus Diseases of Animals, Boddenblick 5a, D-17498 Insel
Riems, Germany. Phone: 49-38351-7266. Fax: 49-38351-7151. E-mail:
klaus.osterrieder{at}rie.bfav.de.
Journal of Virology, December 2001, p. 11307-11318, Vol. 75, No. 23
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.23.11307-11318.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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