The overall goals of the program are to identify the viral and cellular factors involved in the establishment, maintenance and reactivation of herpes simplex virus type 1 latency, and to determine the roles of these factors in the latency process. For this purpose, the following three projects are proposed: Project 1 (D. Knipe) will utilize in situ detection methods to define the nature of the block in immediate-early gene expression during the establishment of latency. Efforts will also be made to identify the type(s) of neurons able to support latency. Viral gene expression in ganglia latently infected with mutants defective in several DNA replication genes will be examined to assess the roles of individual DNA replication genes on gene expression during the establishment and reactivation of latency. The effect of the immune response on gene expression, virus replication and virus spread will be examined by histochemical techniques. Project 2 (D. Coen) will entail the development and use of sensitive PCR procedures to quantify different configurations of viral DNA in latently infected ganglia. Ganglia latently infected with defined mutants will be examined for levels and configurations of viral DNA and compared with viral gene expression by the same mutants (as determined in Projects 1 and 3). Quantitative RNA PCR and cDNA cloning will be used to characterize rare HSV transcripts during the establishment of latency by thymidine kinase (TK) mutants. Ganglia latently infected with mutants defective in TK and ribonucleotide reductase will be examined by in situ hybridization and PCR to determine at what level viral gene expression is blocked in reactivation of such mutants. The roles of individual functional properties of TK and viral DNA polymerase in replication, reactivation and pathogenesis in the peripheral and central nervous systems will be compared. Project 3 (P. Schaffer). Based on the observations that ICPO and the latency-associated transcripts (LATs) are able to stimulate HSV gene expression, and are also required for efficient reactivation from latency, the regions of these two genes that specify transactivating functions involved in reactivation in vivo will be identified and fine-mapped by standard genetic procedures. Efforts will also be made to clone and characterize the genes that specify the recently-identified cellular activating function(s) able to substitute for ICPO during productive infection and able to stimulate viral IE promoters. Collectively, the results of these studies will provide new information concerning the roles of viral and cellular regulatory proteins, and of viral DNA replication proteins in the establishment and reactivation of latency.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program Projects (P01)
Project #
5P01AI024010-07
Application #
3091760
Study Section
Microbiology and Infectious Diseases B Subcommittee (MID)
Project Start
1987-04-01
Project End
1996-03-31
Budget Start
1993-04-01
Budget End
1994-03-31
Support Year
7
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Dana-Farber Cancer Institute
Department
Type
DUNS #
149617367
City
Boston
State
MA
Country
United States
Zip Code
02215
Chen, Shih-Heng; Pearson, Angela; Coen, Donald M et al. (2004) Failure of thymidine kinase-negative herpes simplex virus to reactivate from latency following efficient establishment. J Virol 78:520-3
Chen, S H; Cook, W J; Grove, K L et al. (1998) Human thymidine kinase can functionally replace herpes simplex virus type 1 thymidine kinase for viral replication in mouse sensory ganglia and reactivation from latency upon explant. J Virol 72:6710-5
Pelosi, E; Rozenberg, F; Coen, D M et al. (1998) A herpes simplex virus DNA polymerase mutation that specifically attenuates neurovirulence in mice. Virology 252:364-72
Horsburgh, B C; Chen, S H; Hu, A et al. (1998) Recurrent acyclovir-resistant herpes simplex in an immunocompromised patient: can strain differences compensate for loss of thymidine kinase in pathogenesis? J Infect Dis 178:618-25
Kramer, M F; Chen, S H; Knipe, D M et al. (1998) Accumulation of viral transcripts and DNA during establishment of latency by herpes simplex virus. J Virol 72:1177-85
Pelosi, E; Mulamba, G B; Coen, D M (1998) Penciclovir and pathogenesis phenotypes of drug-resistant Herpes simplex virus mutants. Antiviral Res 37:17-28
Chen, S H; Kramer, M F; Schaffer, P A et al. (1997) A viral function represses accumulation of transcripts from productive-cycle genes in mouse ganglia latently infected with herpes simplex virus. J Virol 71:5878-84
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
Coen, D M (1996) Antiviral drug resistance in herpes simplex virus. Adv Exp Med Biol 394:49-57
Kanangat, S; Babu, J S; Knipe, D M et al. (1996) HSV-1-mediated modulation of cytokine gene expression in a permissive cell line: selective upregulation of IL-6 gene expression. Virology 219:295-300

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