The focus of this project will be to identify the viral cis elements and trans factors which determine HCMV tissue-specific expression. To accomplish this goal, the investigator proposes to characterize further mouse transgenic lines with three separate transgenes, MIEP-b gal (major immediate-early promoter attached to the b-galactosidase gene), a truncated form of the promoter (tMIEP-b gal) and another IE promoter, US3IEb-gal, for tissue specific expression by double labeling procedures. The author also proposes to characterize the tissue-specific expression of transgenic lines containing the 72 kDa autoregulatory IE protein (MIEP 72 kDa) as well as to establish transgenic mice with the other autoregulatory IE isoforms (MIEP 86 kDa and MIEP 55 kDa). Transgenic mice will be established with a putative repressor late protein (MIEP 40 kDa or late promoter [LP] 40 kDa) with an overlapping reading frame with the IE isoforms. Phenotypes associated with expression of these proteins, as well as the effects of these viral proteins on MIEP tissue-specific expression, will be determined. The author also proposes to cross b-gal and IE/late HCMV transgenics to examine the potential of these IE/late proteins in activation or repression of MIEP in tissue. In the final part of this project, the author will try to examine potential scenarios involved in activation of MIEP in normally non-expressing cells. Replication of HCMV in monocyte/macrophages, a reservoir and vector for the virus, is linked to differentiation of the cells. The investigator will utilize peripheral blood monocytes from HCMV transgenic mice to identify differentiation events which may activate the promoter.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA050151-07
Application #
2093622
Study Section
AIDS and Related Research Study Section 3 (ARRC)
Project Start
1989-03-17
Project End
1996-07-31
Budget Start
1995-08-01
Budget End
1996-07-31
Support Year
7
Fiscal Year
1995
Total Cost
Indirect Cost
Name
Oregon Health and Science University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
009584210
City
Portland
State
OR
Country
United States
Zip Code
97239
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Baskar, J F; Smith, P P; Ciment, G S et al. (1996) Developmental analysis of the cytomegalovirus enhancer in transgenic animals. J Virol 70:3215-26
Fish, K N; Stenglein, S G; Ibanez, C et al. (1995) Cytomegalovirus persistence in macrophages and endothelial cells. Scand J Infect Dis Suppl 99:34-40
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Jupp, R; Hoffmann, S; Stenberg, R M et al. (1993) Human cytomegalovirus IE86 protein interacts with promoter-bound TATA-binding protein via a specific region distinct from the autorepression domain. J Virol 67:7539-46
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Ghazal, P; Young, J; Giulietti, E et al. (1991) A discrete cis element in the human immunodeficiency virus long terminal repeat mediates synergistic trans activation by cytomegalovirus immediate-early proteins. J Virol 65:6735-42