Abstract

Herpes simplex virus gene expression is coordinately regulated and temporally controlled. The biosynthesis of early gene transcripts, eg the mRNA for thymidine kinase, is only catalyzed by host RNA polymerase after synthesis of functional immediate early gene products. When cloned immediate early gene are introduced into Hela cells along with a cloned tk gene, tk mRNA is faithfully transcribed. In the absence of immediate early gene sequences we detect little tk specific RNA. The experiments described in this grant application propose to use transient expression assays to define the immediate early genes that act in trans to activate transcription of tk gene. In other studies the sequences that code for the immediate early gene products will be modified in vitro and the effects of these mutations will be studied in a transient expression assay system.
Our aim i s to identify regulatory proteins, their functional domains and to determine if they interact with specific sequences in the virus chromosome.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA017477-12
Application #
3164715
Study Section
Virology Study Section (VR)
Project Start
1985-05-01
Project End
1990-04-30
Budget Start
1986-05-01
Budget End
1987-04-30
Support Year
12
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
Columbia University (N.Y.)
Department
Type
Schools of Medicine
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10027

  Publications

Panagiotidis, C A; Artandi, S; Calame, K et al. (1995) Polyamines alter sequence-specific DNA-protein interactions. Nucleic Acids Res 23:1800-9
Bohenzky, R A; Lagunoff, M; Roizman, B et al. (1995) Two overlapping transcription units which extend across the L-S junction of herpes simplex virus type 1. J Virol 69:2889-97
Burkholder, W F; Panagiotidis, C A; Silverstein, S J et al. (1994) Isolation and characterization of an Escherichia coli DnaK mutant with impaired ATPase activity. J Mol Biol 242:364-77
Bohenzky, R A; Papavassiliou, A G; Gelman, I H et al. (1993) Identification of a promoter mapping within the reiterated sequences that flank the herpes simplex virus type 1 UL region. J Virol 67:632-42
Flanagan, W M; Papavassiliou, A G; Rice, M et al. (1991) Analysis of the herpes simplex virus type 1 promoter controlling the expression of UL38, a true late gene involved in capsid assembly. J Virol 65:769-86
Kemp, L M; Gelman, I H; Silverstein, S J et al. (1990) Regulation of herpes simplex virus immediate-early gene promoters in mouse neuroblastoma cells. Neurosci Lett 118:185-8
Papavassiliou, A G; Silverstein, S J (1990) Interaction of cell and virus proteins with DNA sequences encompassing the promoter/regulatory and leader regions of the herpes simplex virus thymidine kinase gene. J Biol Chem 265:9402-12
Papavassiliou, A G; Silverstein, S J (1990) Characterization of DNA-protein complex formation in nuclear extracts with a sequence from the herpes simplex virus thymidine kinase gene. J Biol Chem 265:1648-57
Zhu, X X; Young, C S; Silverstein, S (1988) Adenovirus vector expressing functional herpes simplex virus ICP0. J Virol 62:4544-53
Gelman, I H; Silverstein, S (1987) Dissection of immediate-early gene promoters from herpes simplex virus: sequences that respond to the virus transcriptional activators. J Virol 61:3167-72

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