Herpes Simplex Virus type l (HSV-l) is a ubiquitous pathogen which causes acute and latent infections in the majority of the population of the United States. The lytic cycle of HSV is the result of a coordinated temporal cascade of virus-specific protein synthesis following entry of the viral DNA into the host cell nucleus. The HSV major DNA-binding protein, commonly designated ICP8, is required for or involved in replication of the viral DNA and regulation of viral gene expression. in carrying out these functions ICP8 interacts with a variety of virus-coded and cellular macromolecules. The overall aim of this proposal is to identify and characterize functional domains within ICP8.
The specific aims to accomplish this purpose are as follows: l. Mapping of the ICP8 zinc-binding domain and determination of the role of zinc in the functions of ICP8. 2. Mapping of the DNA binding-site of ICP8 and identification of specific residues involved in this interaction. 3. Identification and mapping of domains required for cooperative binding of ICP8 to nucleic acids and/or involved in conformational changes within the protein. 4. Identification and functional analysis of regions of ICP8 required for the origin-dependent synthesis of HSV DNA.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI022468-13
Application #
2003342
Study Section
Experimental Virology Study Section (EVR)
Project Start
1985-09-30
Project End
1998-12-31
Budget Start
1997-01-01
Budget End
1997-12-31
Support Year
13
Fiscal Year
1997
Total Cost
Indirect Cost
Name
State University of New York at Buffalo
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
038633251
City
Buffalo
State
NY
Country
United States
Zip Code
14260
Dudas, K C; Scouten, S K; Ruyechan, W T (2001) Conformational change in the herpes simplex single-strand binding protein induced by DNA. Biochem Biophys Res Commun 288:184-90
Dudas, K C; Ruyechan, W T (1998) Identification of a region of the herpes simplex virus single-stranded DNA-binding protein involved in cooperative binding. J Virol 72:257-65
Shelton, L S; Albright, A G; Ruyechan, W T et al. (1994) Retention of the herpes simplex virus type 1 (HSV-1) UL37 protein on single-stranded DNA columns requires the HSV-1 ICP8 protein. J Virol 68:521-5
Ruyechan, W T; Olson, J W (1992) Surface lysine and tyrosine residues are required for interaction of the major herpes simplex virus type 1 DNA-binding protein with single-stranded DNA. J Virol 66:6273-9
Hay, J; Ruyechan, W T (1992) Regulation of herpes simplex virus type 1 gene expression. Curr Top Microbiol Immunol 179:1-14
Gupte, S S; Olson, J W; Ruyechan, W T (1991) The major herpes simplex virus type-1 DNA-binding protein is a zinc metalloprotein. J Biol Chem 266:11413-6
Haffey, M L; Stevens, J T; Terry, B J et al. (1988) Expression of herpes simplex virus type 1 DNA polymerase in Saccharomyces cerevisiae and detection of virus-specific enzyme activity in cell-free lysates. J Virol 62:4493-8
Ruyechan, W T (1988) N-ethylmaleimide inhibition of the DNA-binding activity of the herpes simplex virus type 1 major DNA-binding protein. J Virol 62:810-7
Holmes, A M; Wietstock, S M; Ruyechan, W T (1988) Identification and characterization of a DNA primase activity present in herpes simplex virus type 1-infected HeLa cells. J Virol 62:1038-45
Ruyechan, W T; Chytil, A; Fisher, C M (1986) In vitro characterization of a thermolabile herpes simplex virus DNA-binding protein. J Virol 59:31-6