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Journal of Virology, September 2005, p. 10952-10967, Vol. 79, No. 17
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.17.10952-10967.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Envelope Glycoprotein gB of Kaposi's Sarcoma-Associated Herpesvirus Is Essential for Egress from Infected Cells

Harinivas H. Krishnan,1 Neelam Sharma-Walia,1,3 Ling Zeng,1 Shou-Jiang Gao,2 and Bala Chandran1,3*

Department of Microbiology, Molecular Genetics and Immunology, The University of Kansas Medical Center, Kansas City, Kansas,1 Department of Pediatrics, The University of Texas Health Science Center at San Antonio San Antonio, Texas,2 Department of Microbiology and Immunology, Chicago Medical School, RFUMS, North Chicago, Illinois3

Received 7 February 2005/ Accepted 26 May 2005

Kaposi's sarcoma-associated herpesvirus (KSHV) envelope glycoprotein gB interacts with cell surface heparan sulfate (HS) and {alpha}3ß1 integrin and plays roles in the initial binding and entry into the target cells and in the induction of preexisting host cell signal pathways. To define gB function further, using a bacterial artificial chromosome (BAC) system carrying the KSHV genome (BAC36wt-KSHV), we constructed a recombinant virus genome with the gB open reading frame (ORF) deleted by replacing a 2-kb gB ORF with a 1.3-kb Kanr gene. Stable 293T cells carrying BAC36wt-KSHV and {Delta}gBBAC36-KSHV genomes were generated. Transcript analyses and immunoprecipitation reactions confirmed the absence of gB in the 293T-{Delta}gBBAC36 cells. When monolayers of 293T-BAC36wt and 293T-{Delta}gBBAC36 cells were induced with tetradecanoylphorbol-13-acetate, infectious virus was detected only from the 293T-BAC36wt cell supernatants. No significant amount of DNase I-resistant viral DNA was detected in the supernatants of 293T-{Delta}gBBAC36 cells. BAC36wt-KSHV infected the target cells, and in contrast, no viral DNA and transcripts could be detected in cells infected with {Delta}gBBAC36-KSHV. Electron microscopy of 293T-{Delta}gBBAC36 cells revealed capsids in the nuclei, cytoplasmic vesicles with core-containing capsids, and occasional enveloped virions in the cytoplasm. However, enveloped virus particles were observed in the extracellular compartments of 293T-BAC36wt cells only and not in 293T-{Delta}gBBAC36 cells. Transfection of 293T-{Delta}gBBAC36 cells with plasmid expressing full-length gB restored the recovery of infectious KSHV in the supernatant. These results suggest that, besides its role in virus binding and entry into the target cells, KSHV gB also plays a role in the maturation and egress of virus from the infected cells.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Chicago Medical School, 3333 Green Bay Rd., North Chicago, IL 60064. Phone: (847) 578-8822. Fax: (847) 578-3349. E-mail: bala.chandran{at}rosalindfranklin.edu.

Journal of Virology, September 2005, p. 10952-10967, Vol. 79, No. 17
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.17.10952-10967.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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