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Journal of Virology, March 2008, p. 2802-2812, Vol. 82, No. 6
0022-538X/08/$08.00+0     doi:10.1128/JVI.01550-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

UL74 of Human Cytomegalovirus Contributes to Virus Release by Promoting Secondary Envelopment of Virions{triangledown} ,{dagger}

Xiao Jing Jiang,1,5 Barbara Adler,3 Kerstin Laib Sampaio,1 Margarete Digel,1 Gerhard Jahn,1 Nicole Ettischer,1 York-Dieter Stierhof,2 Laura Scrivano,3 Ulrich Koszinowski,3 Michael Mach,4 and Christian Sinzger1*

Institut für Medizinische Virologie,1 Zentrum für Molekularbiologie der Pflanzen, Eberhard Karls Universität, Tübingen, Germany,2 Max von Pettenkofer Institut für Virologie, Ludwig Maximilians Universität, München, Germany,3 Institut für Klinische und Molekulare Virologie, Universitätsklinikum, Erlangen, Germany,4 Department of Infectious Diseases, Wuhan General Hospital, Wuhan, China5

Received 17 July 2007/ Accepted 20 December 2007

The glycoprotein (g) complex gH/gL represents an essential part of the herpesvirus fusion machinery mediating entry of cell-free virions and cell-associated viral spread. In some herpesviruses additional proteins are associated with gH/gL contributing to the cell tropism of the respective virus. Human cytomegalovirus (HCMV) gH/gL forms complexes with either gO (UL74) or proteins of the UL128-131A gene locus. While a contribution of UL128-131A to endothelial cell tropism is known, the role of gO is less clear. We studied the role of gH/gL-associated proteins in HCMV replication in human foreskin fibroblasts (HFF) and human umbilical vein endothelial cells (HUVEC). Deletions of UL74 alone or in combination with mutations of the UL128-131A gene region were introduced into bacterial artificial chromosome vectors derived from the endotheliotropic strain TB40/E. Deletion of UL74 caused a profound defect regarding virus release from infected HFF and HUVEC. Large numbers of capsids accumulated in the cytoplasm of infected HFF but failed to acquire an envelope. Clear cell type differences were observed in the cell-associated spread of the UL74-defective virus. In HFF, focal growth was severely impaired, whereas it was normal in HUVEC. Deletion of UL131A abolished focal growth in endothelial cells. UL74/UL128-131A dual mutants showed severely impaired reconstitution efficiency. Our data suggest that gO plays a critical role in secondary envelopment and release of cell-free virions independent of the cell type but affects cell-associated growth specifically in HFF, whereas UL128-131A contributes to cell-associated spread in HFF and HUVEC.

* Corresponding author. Mailing address: Institut für Medizinische Virologie, Eberhard Karls Universität, Tübingen, Elfriede Aulhorn Straβe 6, D-72076 Tübingen, Germany. Phone: 7071 2987459. Fax: 7071 295790. E-mail: christian.sinzger{at}med.uni-tuebingen.de

{triangledown} Published ahead of print on 9 January 2008.

{dagger} Supplemental material for this article may be found at http://jvi.asm.org/.

Journal of Virology, March 2008, p. 2802-2812, Vol. 82, No. 6
0022-538X/08/$08.00+0     doi:10.1128/JVI.01550-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Vanarsdall, A. L., Ryckman, B. J., Chase, M. C., Johnson, D. C. (2008). Human Cytomegalovirus Glycoproteins gB and gH/gL Mediate Epithelial Cell-Cell Fusion When Expressed either in cis or in trans. J. Virol. 82: 11837-11850 [Abstract] [Full Text]  
  • Schuessler, A., Sampaio, K. L., Sinzger, C. (2008). Charge Cluster-to-Alanine Scanning of UL128 for Fine Tuning of the Endothelial Cell Tropism of Human Cytomegalovirus. J. Virol. 82: 11239-11246 [Abstract] [Full Text]  
  • Ryckman, B. J., Chase, M. C., Johnson, D. C. (2008). HCMV gH/gL/UL128-131 interferes with virus entry into epithelial cells: Evidence for cell type-specific receptors. Proc. Natl. Acad. Sci. USA 105: 14118-14123 [Abstract] [Full Text]  


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