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Journal of Virology, October 2007, p. 10575-10587, Vol. 81, No. 19
0022-538X/07/$08.00+0     doi:10.1128/JVI.01065-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Horizontal Transmission of Marek's Disease Virus Requires US2, the UL13 Protein Kinase, and gC{triangledown}

Keith W. Jarosinski,1* Neil G. Margulis,1 Jeremy P. Kamil,1 Stephen J. Spatz,2 Venugopal K. Nair,3 and Nikolaus Osterrieder1

Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York,1 Southeast Poultry Research Laboratory, Agriculture Research Service, USDA, Athens, Georgia,2 Viral Oncogenesis Group, Institute for Animal Health, Compton, Berkshire, United Kingdom3

Received 17 May 2007/ Accepted 10 July 2007

Marek's disease virus (MDV) causes a general malaise in chickens that is mostly characterized by the development of lymphoblastoid tumors in multiple organs. The use of bacterial artificial chromosomes (BACs) for cloning and manipulation of the MDV genome has facilitated characterization of specific genes and genomic regions. The development of most MDV BACs, including pRB-1B-5, derived from a very virulent MDV strain, involved replacement of the US2 gene with mini-F vector sequences. However, when reconstituted viruses based on pRB-1B were used in pathogenicity studies, it was discovered that contact chickens housed together with experimentally infected chickens did not contract Marek's disease (MD), indicating a lack of horizontal transmission. Staining of feather follicle epithelial cells in the skins of infected chickens showed that virus was present but was unable to be released and/or infect susceptible chickens. Restoration of US2 and removal of mini-F sequences within viral RB-1B did not alter this characteristic, although in vivo viremia levels were increased significantly. Sequence analyses of pRB-1B revealed that the UL13, UL44, and US6 genes encoding the UL13 serine/threonine protein kinase, glycoprotein C (gC), and gD, respectively, harbored frameshift mutations. These mutations were repaired individually, or in combination, using two-step Red mutagenesis. Reconstituted viruses were tested for replication, MD incidence, and their abilities to horizontally spread to contact chickens. The experiments clearly showed that US2, UL13, and gC in combination are essential for horizontal transmission of MDV and that none of the genes alone is able to restore this phenotype.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853. Phone: (607) 253-4067. Fax: (607) 253-3384. E-mail: kwj4{at}cornell.edu

{triangledown} Published ahead of print on 18 July 2007.

Journal of Virology, October 2007, p. 10575-10587, Vol. 81, No. 19
0022-538X/07/$08.00+0     doi:10.1128/JVI.01065-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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