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Journal of Virology, June 2004, p. 6389-6398, Vol. 78, No. 12
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.12.6389-6398.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Kaposi's Sarcoma-Associated Herpesvirus Glycoprotein K8.1 Is Dispensable for Virus Entry

Rafael E. Luna,1,2 Fuchun Zhou,3,4 Abolgashem Baghian,1 Vladimir Chouljenko,1 Bagher Forghani,5 Shou-Jiang Gao,3,4,6,7 and Konstantin G. Kousoulas1*

Division of Biotechnology and Molecular Medicine, School of Veterinary Medicine,1 Department of Biological Sciences, Louisiana State University, Baton Rouge, Louisiana 70803,2 Tumor Virology Program, Children's Cancer Research Institute,3 Departments of Pediatrics,4 Microbiology and Immunology,6 Medicine, The University of Texas Health Science Center at San Antonio, San Antonio, Texas 78229,7 Viral Immunology Section, Viral and Rickettsial Disease Laboratory, California Department of Health Services, Richmond, California 948045

Received 9 December 2003/ Accepted 5 February 2004

Kaposi's sarcoma-associated herpesvirus (KSHV) is considered the etiologic agent of Kaposi's sarcoma and several lymphoproliferative disorders. Recently, the KSHV genome was cloned into a bacterial artificial chromosome and used to construct a recombinant KSHV carrying a deletion of the viral interferon regulatory factor gene (F. C. Zhou, Y. J. Zhang, J. H. Deng, X. P. Wang, H. Y. Pan, E. Hettler, and S. J. Gao, J. Virol. 76:6185-6196, 2002). The K8.1 glycoprotein is a structural component of the KSHV particle and is thought to facilitate virus entry by binding to heparan sulfate moieties on cell surfaces. To further address the role of K8.1 in virus infectivity, a K8.1-null recombinant virus (BAC36{Delta}K8.1) was constructed by deletion of most of the K8.1 open reading frame and insertion of a kanamycin resistance gene cassette within the K8.1 gene. Southern blotting and diagnostic PCR confirmed the presence of the engineered K8.1 gene deletion. Transfection of the wild-type genome (BAC36) and mutant genome (BAC36{Delta}K8.1) DNAs into 293 cells in the presence or absence of the complementing plasmid (pCDNAK8.1A), transiently expressing the K8.1A gene, produced infectious virions in the supernatants of transfected cells. These results demonstrated that the K8.1 glycoprotein is not required for KSHV entry into 293 cells.

* Corresponding author. Mailing address: BIOMMED, School of Veterinary Medicine, Louisiana State University, Skip Bertman Dr., Baton Rouge, LA 70803. Phone: (225) 578-9683. Fax: (225) 578-9655. E-mail: vtgusk{at}lsu.edu.

Journal of Virology, June 2004, p. 6389-6398, Vol. 78, No. 12
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.12.6389-6398.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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