Abstract

Herpes simplex virus (HSV) is the etiological agent of a number of diseases including herpes labialis and there is a correlation between HSV infections and the development of certain neoplastic diseases. HSV induces approximately 50 polypeptides in permissively infected cells and some of these possess enzymatic activity. However, except for the HSV induced DNA polymerase and thymidine kinase, little is known concerning the roles of other HSV-induced enzymes in replication or in antiviral chemotherapy. The goal of this research is to determine whether the HSV induced deoxyuridine triphosphate nucleotidohydrolase (dUTPase, EC 2.6.1.23) can be used as a target site for the development of specific antiviral compounds and to determine what role this enzyme may have in regulating the chemotherapeutic effectiveness of specific antiviral agents. To accomplish this, we plan to use purified HSV induced and cellular dUTPases to elucidate features concerning the active-site of the dUTPases, to determine differences in the steric-binding properties of these enzymes and to determine the mechanism(s) by which these dUTPases are inhibited by various mercury (II) compounds. We will construct, using insertional and site-directed mutagenesis HSV mutants that are defective in dUTPase activity and that have altered dUTPase activity. These mutants as well as wild-type virus will be used in in vivo studies to determine how changes in deoxyuridine (dUrd) pools will effect incorporation of dUTP into HSV DNA and to determine how incorporation of dUTP into the DNA effects HSV replication.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
2R01DE006866-04A2
Application #
3220316
Study Section
Experimental Therapeutics Subcommittee 2 (ET)
Project Start
1984-03-01
Project End
1991-06-30
Budget Start
1988-07-01
Budget End
1989-06-30
Support Year
4
Fiscal Year
1988
Total Cost
Indirect Cost

  Institution

Name
Ohio State University
Department
Type
Schools of Medicine
DUNS #
098987217
City
Columbus
State
OH
Country
United States
Zip Code
43210

  Publications

Waldman, W J; Roberts, W H; Davis, D H et al. (1991) Preservation of natural endothelial cytopathogenicity of cytomegalovirus by propagation in endothelial cells. Arch Virol 117:143-64
Tsai, C H; Williams, M V; Glaser, R (1991) Characterization of two monoclonal antibodies to Epstein-Barr virus diffuse early antigen which react to two different epitopes and have different biological function. J Virol Methods 33:47-52
Holliday, J; Williams, M V (1991) Inhibition of herpes simplex virus types 1 and 2 replication in vitro by mercurithio analogs of deoxyuridine. Antiviral Res 16:197-203
Winters, T A; Williams, M V (1990) Use of the PBS2 uracil-DNA glycosylase inhibitor to differentiate the uracil-DNA glycosylase activities encoded by herpes simplex virus types 1 and 2. J Virol Methods 29:233-42
Tsai, C H; Williams, M V; Glaser, R (1990) A monoclonal antibody that neutralizes Epstein-Barr virus, human cytomegalovirus, human herpesvirus 6, and bacteriophage T4 DNA polymerases. Proc Natl Acad Sci U S A 87:7963-7
Tsai, C H; Williams, M V; Glaser, R (1990) A monoclonal antibody that neutralizes Epstein-Barr virus DNA polymerase activity. Intervirology 31:215-22
Williams, M V; Pollack, J D (1990) A mollicute (mycoplasma) DNA repair enzyme: purification and characterization of uracil-DNA glycosylase. J Bacteriol 172:2979-85
Williams, M V; Ablashi, D V; Salahuddin, S Z et al. (1989) Demonstration of the human herpesvirus 6-induced DNA polymerase and DNase. Virology 173:223-30
Williams, M V (1988) Herpes simplex virus-induced dUTPase: target site for antiviral chemotherapy. Virology 166:262-4
Williams, M V; Boak, A; Glaser, R (1988) Antigenic variation in alkaline deoxyribonuclease induced by three different strains of Epstein-Barr virus. J Med Virol 26:207-15

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