The human herpesviruses cause a spectrum of clinically significant acute and life-long latent infections, and some can transform cells morphologically and oncogenically. During productive infection, all or nearly all viral genes are expressed, resulting in the synthesis of new infectious virus, cell death, and clinical disease. A better understanding of the mechanisms involved in the regulation of viral gene expression during productive infection should shed light on the disease-producing properties of herpesviruses and may ultimately provide novel approaches to intervention in the herpesvirus life cycle. Immediate-early (IE) viral proteins have been implicated as primary factors in regulating viral gene expression during productive infection. In order to define the functions of four of the five HSV IE proteins, a series of small in-frame mutations will be introduced into coding sequences of the genes specifying ICPs O and 27. The effects of these mutations on the transcriptional and translational programs of the mutants will be assessed. The proteins themselves will be purified and characterized for their abilities 1) to bind to specific DNA sequences in viral promoter, 2) to regulate viral gene expression in an in vitro transcription assay, and 3) to bind to other viral and cellular proteins. The DNA sequences in viral promoters that respond to the regulatory activities of ICPs 0 and 27 will be identified by systematic mutational and functional analysis. Mutations will also be introduced into coding sequences of the genes specifying ICPs 22 and 47, and the effects of splicing on the expression of the genes specifying ICPs 0, 22 and 47 will be investigated by substituting unspliced cDNA copies of these genes for their spliced counterparts in the viral genome. The resulting mutant viruses will be characterized with regard to the parameters of viral gene expression. Taken together, these studies should provide a better understanding of the functions of HSSV-1 IE proteins and the manner in which they mediate viral gene expression.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37CA020260-18
Application #
2086816
Study Section
Experimental Virology Study Section (EVR)
Project Start
1980-03-01
Project End
1994-12-31
Budget Start
1994-01-01
Budget End
1994-12-31
Support Year
18
Fiscal Year
1994
Total Cost
Indirect Cost
Name
Dana-Farber Cancer Institute
Department
Type
DUNS #
149617367
City
Boston
State
MA
Country
United States
Zip Code
02215
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Jordan, R; Pepe, J; Schaffer, P A (1998) Characterization of a nerve growth factor-inducible cellular activity that enhances herpes simplex virus type 1 gene expression and replication of an ICP0 null mutant in cells of neural lineage. J Virol 72:5373-82
Lee, L Y; Schaffer, P A (1998) A virus with a mutation in the ICP4-binding site in the L/ST promoter of herpes simplex virus type 1, but not a virus with a mutation in open reading frame P, exhibits cell-type-specific expression of gamma(1)34.5 transcripts and latency-associated transc J Virol 72:4250-64
Schang, L M; Phillips, J; Schaffer, P A (1998) Requirement for cellular cyclin-dependent kinases in herpes simplex virus replication and transcription. J Virol 72:5626-37
Jordan, R; Schaffer, P A (1997) Activation of gene expression by herpes simplex virus type 1 ICP0 occurs at the level of mRNA synthesis. J Virol 71:6850-62
Frazier, D P; Cox, D; Godshalk, E M et al. (1996) The herpes simplex virus type 1 latency-associated transcript promoter is activated through Ras and Raf by nerve growth factor and sodium butyrate in PC12 cells. J Virol 70:7424-32
Wu, N; Watkins, S C; Schaffer, P A et al. (1996) Prolonged gene expression and cell survival after infection by a herpes simplex virus mutant defective in the immediate-early genes encoding ICP4, ICP27, and ICP22. J Virol 70:6358-69
Zhu, Z; DeLuca, N A; Schaffer, P A (1996) Overexpression of the herpes simplex virus type 1 immediate-early regulatory protein, ICP27, is responsible for the aberrant localization of ICP0 and mutant forms of ICP4 in ICP4 mutant virus-infected cells. J Virol 70:5346-56
Frazier, D P; Cox, D; Godshalk, E M et al. (1996) Identification of cis-acting sequences in the promoter of the herpes simplex virus type 1 latency-associated transcripts required for activation by nerve growth factor and sodium butyrate in PC12 cells. J Virol 70:7433-44
Yao, F; Schaffer, P A (1995) An activity specified by the osteosarcoma line U2OS can substitute functionally for ICP0, a major regulatory protein of herpes simplex virus type 1. J Virol 69:6249-58

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