Herpes simplex virus (HSV) infection often results in a recurrent disease that manifests as skin lesions in the facial or genital area. Herpes virus infections are a leading cause of blindness and fatal endemic enecphalitis. Reactivation of herpes is a serious problem for individuals with immune systems compromised by chemotherapy or HIV infection. Productive infection and the consequent cell destruction by herpes virus requires activation of viral gene expression by a viral protein designated ICP4 (IE175, Vmw175). Initial characterization of the structure and function of ICP4 suggests that it is an ideal target for anti-viral drugs. ICP4 has been conceptually divided into 5 regions on the basis of comparative sequence analysis with related members of the alphaherpesvirus family. These proteins share extensive homology in regions 2 and 4. There is evidence for a transactivation domain in region 1, a DNA binding domain in region 2, and nuclear localization signal in region 3. Although all 5 domains are required for full ICP4 activity, no specific functions have been assigned to regions 4 and 5. The long-term goal of this investigation is to define the structural basis for the functions of ICP4.
The specific aims of this project are (1) to characterize the ICP4 during productive infection; and (4) to investigate interactions between ICP4 and a component of the 20S proteasome. The roles of TAD-1 in productive infection and pathogenesis will be determined by infection of cells and animals with a TAD-1 negative mutant. TAD-1 will be further defined by analysis of the effect of point mutations in TAD-1 on transactivation. Coimmunoprecipitation assays with mutant proteins will be performed to define the residues in region 4 that are responsible for TBP/TFIIB interactions. The biological significance of the TBP/TFIIB binding sites in region 4 will be investigated by complementation assays and analysis of viral mutants. Interactions between ICP4 and proteasomes will be investigated in transfected and infected cells. The results of these analysis will be integrated into a working model for ICP4.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI017246-17
Application #
2886387
Study Section
Experimental Virology Study Section (EVR)
Program Officer
Beisel, Christopher E
Project Start
1980-07-01
Project End
2002-03-31
Budget Start
1999-04-01
Budget End
2000-03-31
Support Year
17
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Medical College of Wisconsin
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
073134603
City
Milwaukee
State
WI
Country
United States
Zip Code
53226
Wilcox, Kent W; Sheriff, Scott; Isaac, Anne et al. (2005) SP100B is a repressor of gene expression. J Cell Biochem 95:352-65
Bruce, James W; Wilcox, Kent W (2002) Identification of a motif in the C terminus of herpes simplex virus regulatory protein ICP4 that contributes to activation of transcription. J Virol 76:195-207
Taylor, J L; Unverrich, D; O'Brien, W J et al. (2000) Interferon coordinately inhibits the disruption of PML-positive ND10 and immediate-early gene expression by herpes simplex virus. J Interferon Cytokine Res 20:805-15
Xiao, W; Pizer, L I; Wilcox, K W (1997) Identification of a promoter-specific transactivation domain in the herpes simplex virus regulatory protein ICP4. J Virol 71:1757-65
Wu, C L; Wilcox, K W (1991) The conserved DNA-binding domains encoded by the herpes simplex virus type 1 ICP4, pseudorabies virus IE180, and varicella-zoster virus ORF62 genes recognize similar sites in the corresponding promoters. J Virol 65:1149-59
Wu, C L; Wilcox, K W (1990) Codons 262 to 490 from the herpes simplex virus ICP4 gene are sufficient to encode a sequence-specific DNA binding protein. Nucleic Acids Res 18:531-8
Kattar-Cooley, P; Wilcox, K W (1989) Characterization of the DNA-binding properties of herpes simplex virus regulatory protein ICP4. J Virol 63:696-704
Tedder, D G; Everett, R D; Wilcox, K W et al. (1989) ICP4-binding sites in the promoter and coding regions of the herpes simplex virus gD gene contribute to activation of in vitro transcription by ICP4. J Virol 63:2510-20
Faber, S W; Wilcox, K W (1988) Association of herpes simplex virus regulatory protein ICP4 with sequences spanning the ICP4 gene transcription initiation site. Nucleic Acids Res 16:555-70
Faber, S W; Wilcox, K W (1986) Characterization of a herpes simplex virus regulatory protein: aggregation and phosphorylation of a temperature-sensitive variant of ICP 4. Arch Virol 91:297-312

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