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Virus-Cell Interactions

Two Gamma Interferon-Activated Site-Like Elements in the Human Cytomegalovirus Major Immediate-Early Promoter/Enhancer Are Important for Viral Replication

James Netterwald, Shaojun Yang, Weijia Wang, Salena Ghanny, Michael Cody, Patricia Soteropoulos, Bin Tian, Walter Dunn, Fenyong Liu, Hua Zhu
James Netterwald
1Department of Microbiology and Molecular Genetics
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Shaojun Yang
1Department of Microbiology and Molecular Genetics
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Weijia Wang
1Department of Microbiology and Molecular Genetics
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Salena Ghanny
2Center for Applied Genomics, Public Health Research Institute, International Center for Public Health, Newark, New Jersey
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Michael Cody
2Center for Applied Genomics, Public Health Research Institute, International Center for Public Health, Newark, New Jersey
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Patricia Soteropoulos
1Department of Microbiology and Molecular Genetics
2Center for Applied Genomics, Public Health Research Institute, International Center for Public Health, Newark, New Jersey
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Bin Tian
3Department of Molecular Biology and Biochemistry, New Jersey Medical School, University of Medicine and Dentistry of New Jersey
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Walter Dunn
4Division of Infectious Diseases, School of Public Health, University of California, Berkeley, California
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Fenyong Liu
4Division of Infectious Diseases, School of Public Health, University of California, Berkeley, California
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Hua Zhu
1Department of Microbiology and Molecular Genetics
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  • For correspondence: zhuhu@umdnj.edu
DOI: 10.1128/JVI.79.8.5035-5046.2005
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ABSTRACT

Human cytomegalovirus (HCMV) infection directly initiates a signal transduction pathway that leads to activation of a large number of cellular interferon-stimulated genes (ISGs). Our previous studies demonstrated that two interferon response elements, the interferon-stimulated response element and gamma interferon-activated site (GAS), in the ISG promoters serve as HCMV response sites (VRS). Interestingly, two GAS-like VRS elements (VRS1) were also present in the HCMV major immediate-early promoter-enhancer (MIEP/E). In this study, the importance of these VRS elements in viral replication was investigated. We demonstrate that the expression of the major IE genes, IE1 and IE2, is interferon inducible. To understand the biological significance of this signal transduction pathway in HCMV major IE expression, the two VRS1 in the MIEP/E were mutated. Mutant HCMVs in which the VRS elements were deleted or that contained point mutations grew dramatically more slowly than wild-type virus at a low multiplicity of infection (MOI). Insertion of wild-type VRS1 into the mutant viral genome rescued the slow growth phenotype. Furthermore, the expression levels of major IE RNAs and proteins were greatly reduced during infection with the VRS mutants at a low MOI. HCMV microarray analysis indicated that infection of host cells with the VRS mutant virus resulted in a global reduction in the expression of viral genes. Collectively, these data demonstrate that the two VRS elements in the MIEP/E are necessary for efficient viral gene expression and replication. This study suggests that although the HCMV-initiated signal transduction pathway results in induction of cellular antiviral genes, it also functions to stimulate viral major IE gene expression. This might be a new viral strategy in which the pathway is used to regulate gene expression and play a role in reactivation.

  • Copyright © 2005 American Society for Microbiology
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Two Gamma Interferon-Activated Site-Like Elements in the Human Cytomegalovirus Major Immediate-Early Promoter/Enhancer Are Important for Viral Replication
James Netterwald, Shaojun Yang, Weijia Wang, Salena Ghanny, Michael Cody, Patricia Soteropoulos, Bin Tian, Walter Dunn, Fenyong Liu, Hua Zhu
Journal of Virology Mar 2005, 79 (8) 5035-5046; DOI: 10.1128/JVI.79.8.5035-5046.2005

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Two Gamma Interferon-Activated Site-Like Elements in the Human Cytomegalovirus Major Immediate-Early Promoter/Enhancer Are Important for Viral Replication
James Netterwald, Shaojun Yang, Weijia Wang, Salena Ghanny, Michael Cody, Patricia Soteropoulos, Bin Tian, Walter Dunn, Fenyong Liu, Hua Zhu
Journal of Virology Mar 2005, 79 (8) 5035-5046; DOI: 10.1128/JVI.79.8.5035-5046.2005
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KEYWORDS

cytomegalovirus
Enhancer Elements, Genetic
Genes, Immediate-Early
Interferon-gamma
Promoter Regions, Genetic

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