Human cytomegalovirus (HCMV) can cause congenital neurological damage and disseminated infections resulting in pneumonitis, retinitis, hepatitis, and gastroenteritis. HCMV infections have been associated with accelerated atherosclerosis and with coronary restenosis following angioplasty. HCMV infections in solid organ and bone marrow transplant recipients are a significant cause of morbidity. Both hematopoietic cells of the bone marrow and monocytes of the blood can be latently infected with HCMV. Productive infection occurs in a variety of terminally differentiated cells including fibroblast, cytotrophoblast, smooth muscle, endothelial, epithelial, and mircoglial cells and in macrophages. Abortive infection occurs in polymorphonuclear cells. Our laboratory is interested in viral DNA regulatory elements and in viral and cellular proteins that regulate latency, persistent infection, and productive infection.
In Specific Aim I, we propose to determine the role of specific regulatory elements in the proximal 3'-end of the major immediate early (MIE) enhancer on viral gene expression in undifferentiated cells relevant to HCMV latency. Regulatory elements that function to repress transcription from the MIE promoter upon binding a regulator protein in undifferentiated cells of the myeloid lineage will be mutated in the context of the viral genome, and recombinant viruses will be analyzed.
In Specific Aim II, we propose to characterize a repressor-boundary region 5' to the MIE enhancer that prevents the MIE enhancer from affecting transcription from the flanking UL1 27 promoter. We have identified a repressor-boundary region between the UL127 promoter and the MIE enhancer that is unique to HCMV and to date, not found in other herpesviruses or DNA viruses. Due to its location, the repressor-boundary region has a role in the temporal expression of the immediate early (IE) and early HCMV genes that flank the MIE enhancer. Downstream of the MIE promoter are the MIE genes, IE1 and 1E2, which encode for proteins of 72 (1E72) and 86 kDa (lE86), respectively. These viral proteins are key regulatory proteins for efficient productive infection.
In Specific Aim I ll, we propose to determine early gene regulation in recombinant viruses with both the IEl and IE2 genes deleted and to determine the effects of HCMV tEl and 1E2 gene products on viral and host cell gene expression relevant to the viral life cycle. We will investigate the functions of this replication defective HCMV in activation of viral or cellular gene expression and compare these functions to those of the 1E72 and 1E86 proteins.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI013562-25
Application #
6532657
Study Section
Virology Study Section (VR)
Program Officer
Beisel, Christopher E
Project Start
1976-09-01
Project End
2006-05-31
Budget Start
2002-08-01
Budget End
2003-05-31
Support Year
25
Fiscal Year
2002
Total Cost
$264,600
Indirect Cost
Name
University of Iowa
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
041294109
City
Iowa City
State
IA
Country
United States
Zip Code
52242
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Du, Guixin; Stinski, Mark F (2013) Interaction network of proteins associated with human cytomegalovirus IE2-p86 protein during infection: a proteomic analysis. PLoS One 8:e81583
Isomura, Hiroki; Stinski, Mark F (2013) Coordination of late gene transcription of human cytomegalovirus with viral DNA synthesis: recombinant viruses as potential therapeutic vaccine candidates. Expert Opin Ther Targets 17:157-66
Isomura, Hiroki; Stinski, Mark F; Murata, Takayuki et al. (2011) The human cytomegalovirus gene products essential for late viral gene expression assemble into prereplication complexes before viral DNA replication. J Virol 85:6629-44
Du, Guixin; Dutta, Nirmal; Lashmit, Philip et al. (2011) Alternative splicing of the human cytomegalovirus major immediate-early genes affects infectious-virus replication and control of cellular cyclin-dependent kinase. J Virol 85:804-17
Isomura, Hiroki; Stinski, Mark F; Murata, Takayuki et al. (2010) The human cytomegalovirus UL76 gene regulates the level of expression of the UL77 gene. PLoS One 5:e11901
Lashmit, Philip; Wang, Shuhui; Li, Hongmei et al. (2009) The CREB site in the proximal enhancer is critical for cooperative interaction with the other transcription factor binding sites to enhance transcription of the major intermediate-early genes in human cytomegalovirus-infected cells. J Virol 83:8893-904