Abstract

I propose to analyze the molecular basis of spontaneous mutagenesis in mammalian cells infected with herpes simplex virus type 1. I have chosen this system because herpes virus codes for many DNA metabolic functions (including a DNA polymerase), making it readily susceptible to biochemical and genetic analysis. Also, mutations in the viral thymidine kinase gene are readily detected and characterized. I will test whether the viral DNA polymerase influences the production of point mutations. The approach is to determine whether viral strains carrying altered DNA polymerases exhibit altered production of point mutations in the viral thymidine kinase gene. DNA polymerase from mutator and/or antimutator strains identified on this basis will be purified and compared biochemically to the wildtype polymerase. Specific comparisons will be made of (1) the abilities of these enzymes to select correct base pairs during polymerizarion and (2) the levels and specificities of the polymerase-associated 3 feet to 5 feet exonucleases. Mutations due to genetic rearrangements are also known to be quite common in mammalian cells. Further objectives of this study are to deternine (1) wheteher this class of mutations occurs in the herpes thymidine kinase gene, (2) if so, what types of rearrangements are observed and (3) whether tranposable elements play a role in mutant production.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM032646-03
Application #
3281684
Study Section
Virology Study Section (VR)
Project Start
1983-04-01
Project End
1986-03-31
Budget Start
1985-04-01
Budget End
1986-03-31
Support Year
3
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
University of Arizona
Department
Type
Schools of Arts and Sciences
DUNS #
City
Tucson
State
AZ
Country
United States
Zip Code
85722

  Publications

Wang, Y S; Woodward, S; Hall, J D (1992) Use of suppressor analysis to identify DNA polymerase mutations in herpes simplex virus which affect deoxynucleoside triphosphate substrate specificity. J Virol 66:1814-6
Wang, Y S; Hall, J D (1990) Characterization of a major DNA-binding domain in the herpes simplex virus type 1 DNA-binding protein (ICP8). J Virol 64:2082-9
Hall, J D; Wang, Y S; Pierpont, J et al. (1989) Aphidicolin resistance in herpes simplex virus type I reveals features of the DNA polymerase dNTP binding site. Nucleic Acids Res 17:9231-44
Hall, J D; Woodward, S (1989) Aphidicolin resistance in herpes simplex virus type 1 appears to alter substrate specificity in the DNA polymerase. J Virol 63:2874-6
Hall, J D; Myers, E W (1988) A software tool for finding locally optimal alignments in protein and nucleic acid sequences. Comput Appl Biosci 4:35-40
Hall, J D (1988) Modeling functional sites in DNA polymerases. Trends Genet 4:42-6
Chatterjee, S; Petzold, S J; Berger, S J et al. (1987) Strategy for selection of cell variants deficient in poly(ADP-ribose) polymerase. Exp Cell Res 172:245-57
Hall, J D; Gibbs, J S; Coen, D M et al. (1986) Structural organization and unusual codon usage in the DNA polymerase gene from herpes simplex virus type 1. DNA 5:281-8
Hall, J D; Furman, P A; St Clair, M H et al. (1985) Reduced in vivo mutagenesis by mutant herpes simplex DNA polymerase involves improved nucleotide selection. Proc Natl Acad Sci U S A 82:3889-93
Gibbs, J S; Chiou, H C; Hall, J D et al. (1985) Sequence and mapping analyses of the herpes simplex virus DNA polymerase gene predict a C-terminal substrate binding domain. Proc Natl Acad Sci U S A 82:7969-73

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