This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Chau, N. H.
Right arrow Articles by Kerry, J. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Chau, N. H.
Right arrow Articles by Kerry, J. A.

Next Article 

Journal of Virology, February 1999, p. 863-870, Vol. 73, No. 2
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Transcriptional Regulation of the Human Cytomegalovirus US11 Early Gene

Nha H. Chau, Cynthia D. Vanson, and Julie A. Kerry*

Department of Microbiology and Molecular Cell Biology, Eastern Virginia Medical School, Norfolk, Virginia 23501

Received 9 September 1998/Accepted 22 October 1998

The human cytomegalovirus (HCMV) US11 early gene encodes a protein involved in the down-regulation of major histocompatibility complex class I cell surface expression in HCMV-infected cells. Consequently, this gene is thought to play an important role in HCMV evasion of immune recognition. In this study, we examined the transcriptional regulation of US11 gene expression. Analysis of deletions within the US11 promoter suggests that two sequence elements are important for activation by the viral immediate-early (IE) proteins. Deletion of a CREB site located at -83 relative to the cap site resulted in a reduction in promoter activity to 50% of the wild-type level. Deletion of an additional ATF site immediately upstream of the TATA box resulted in abrogation of responsiveness to the IE proteins. To confirm the role of the CREB and ATF sites within the US11 promoter, mutagenesis of these two sites, both individually and in combination, was carried out. Results indicate that both the CREB element and the ATF site were required for full promoter activity, with the ATF site critical for US11 promoter activation. The loss of transcriptional activation correlated with a loss of cellular proteins binding to the mutated US11 promoter elements. In combination with the viral IE proteins, the HCMV tegument protein pp71 (UL82) was found to up-regulate the US11 promoter by six- to sevenfold in transient assays. These results suggest that pp71 may contribute to the activation of the US11 promoter at early times after infection. Up-regulation by pp71 required the presence of the CREB and ATF sites within the US11 promoter for full activation. The role of the ATF and CREB elements in regulating US11 gene expression during viral infection was then assessed. The US11 gene is not required for replication of HCMV in tissue culture. This property was exploited to generate US11 promoter mutants regulating expression of the endogenous US11 gene in the natural genomic context. We generated recombinant HCMV that contained the US11 promoter with mutations in either the CREB or ATF element or both regulating the expression of the endogenous US11 gene. Northern blot analysis of infected cell mRNA revealed that mutation of the CREB element reduced US11 mRNA expression to approximately 25% of that of the wild-type promoter, with identical kinetics of expression. Mutation of the ATF site alone reduced US11 mRNA levels to 6% of that of the wild-type promoter, with mRNA detectable only at 8 h after infection. Mutation of both the CREB and ATF elements in the US11 promoter reduced US11 gene expression to undetectable levels. These results demonstrate that the CREB and ATF sites cooperate to regulate the US11 promoter in HCMV-infected cells.

* Corresponding author. Mailing address: Department of Microbiology and Molecular Cell Biology, Eastern Virginia Medical School, P.O. Box 1980, 700 W. Olney Rd., Norfolk, VA 23501. Phone: (757) 446-5663. Fax: (757) 624-2255. E-mail: JAK{at}BORG.EVMS.EDU.

Journal of Virology, February 1999, p. 863-870, Vol. 73, No. 2
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Lukashchuk, V., McFarlane, S., Everett, R. D., Preston, C. M. (2008). Human Cytomegalovirus Protein pp71 Displaces the Chromatin-Associated Factor ATRX from Nuclear Domain 10 at Early Stages of Infection. J. Virol. 82: 12543-12554 [Abstract] [Full Text]  
  • Cantrell, S. R., Bresnahan, W. A. (2006). Human Cytomegalovirus (HCMV) UL82 Gene Product (pp71) Relieves hDaxx-Mediated Repression of HCMV Replication.. J. Virol. 80: 6188-6191 [Abstract] [Full Text]  
  • Preston, C. M., Nicholl, M. J. (2006). Role of the cellular protein hDaxx in human cytomegalovirus immediate-early gene expression.. J. Gen. Virol. 87: 1113-1121 [Abstract] [Full Text]  
  • Bego, M., Maciejewski, J., Khaiboullina, S., Pari, G., St. Jeor, S. (2005). Characterization of an Antisense Transcript Spanning the UL81-82 Locus of Human Cytomegalovirus. J. Virol. 79: 11022-11034 [Abstract] [Full Text]  
  • Cantrell, S. R., Bresnahan, W. A. (2005). Interaction between the Human Cytomegalovirus UL82 Gene Product (pp71) and hDaxx Regulates Immediate-Early Gene Expression and Viral Replication. J. Virol. 79: 7792-7802 [Abstract] [Full Text]  
  • Forgacs, E., Gupta, S. K., Kerry, J. A., Semmes, O. J. (2005). The bZIP Transcription Factor ATFx Binds Human T-Cell Leukemia Virus Type 1 (HTLV-1) Tax and Represses HTLV-1 Long Terminal Repeat-Mediated Transcription. J. Virol. 79: 6932-6939 [Abstract] [Full Text]  
  • Preston, C. M., Nicholl, M. J. (2005). Human Cytomegalovirus Tegument Protein pp71 Directs Long-Term Gene Expression from Quiescent Herpes Simplex Virus Genomes. J. Virol. 79: 525-535 [Abstract] [Full Text]  
  • Fang, L., Stevens, J. L., Berk, A. J., Spindler, K. R. (2004). Requirement of Sur2 for Efficient Replication of Mouse Adenovirus Type 1. J. Virol. 78: 12888-12900 [Abstract] [Full Text]  
  • Kronschnabl, M., Stamminger, T. (2003). Synergistic induction of intercellular adhesion molecule-1 by the human cytomegalovirus transactivators IE2p86 and pp71 is mediated via an Sp1-binding site. J. Gen. Virol. 84: 61-73 [Abstract] [Full Text]  
  • Hannenhalli, S., Levy, S. (2002). Predicting transcription factor synergism. Nucleic Acids Res 30: 4278-4284 [Abstract] [Full Text]  
  • Ishov, A. M., Vladimirova, O. V., Maul, G. G. (2002). Daxx-Mediated Accumulation of Human Cytomegalovirus Tegument Protein pp71 at ND10 Facilitates Initiation of Viral Infection at These Nuclear Domains. J. Virol. 76: 7705-7712 [Abstract] [Full Text]  
  • Marshall, K. R., Rowley, K. V., Rinaldi, A., Nicholson, I. P., Ishov, A. M., Maul, G. G., Preston, C. M. (2002). Activity and intracellular localization of the human cytomegalovirus protein pp71. J. Gen. Virol. 83: 1601-1612 [Abstract] [Full Text]  
  • Hofmann, H., Sindre, H., Stamminger, T. (2002). Functional Interaction between the pp71 Protein of Human Cytomegalovirus and the PML-Interacting Protein Human Daxx. J. Virol. 76: 5769-5783 [Abstract] [Full Text]  
  • Gawn, J. M., Greaves, R. F. (2002). Absence of IE1 p72 Protein Function during Low-Multiplicity Infection by Human Cytomegalovirus Results in a Broad Block to Viral Delayed-Early Gene Expression. J. Virol. 76: 4441-4455 [Abstract] [Full Text]  
  • Johnson, R. A., Ma, X.-L., Yurochko, A. D., Huang, E.-S. (2001). The role of MKK1/2 kinase activity in human cytomegalovirus infection. J. Gen. Virol. 82: 493-497 [Abstract] [Full Text]  
  • Bresnahan, W. A., Shenk, T. E. (2000). UL82 virion protein activates expression of immediate early viral genes in human cytomegalovirus-infected cells. Proc. Natl. Acad. Sci. USA 97: 14506-14511 [Abstract] [Full Text]  
  • Johnson, R. A., Huong, S.-M., Huang, E.-S. (2000). Activation of the Mitogen-Activated Protein Kinase p38 by Human Cytomegalovirus Infection through Two Distinct Pathways: a Novel Mechanism for Activation of p38. J. Virol. 74: 1158-1167 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»