The analysis of the regulatory phenomena controlling the coordinately and sequentially ordered synthesis of herpes simplex virus type I (HSV-1) specific proteins during infection continues to be the objective of this research. Two new initiatives are proposed. The first is an extension of previous studies on the regulation of expression of the glycoprotein C (gC) gene, a late gene. Late gene expression requires transactivation by the gene products of two immediate-early genes, ICP4 and ICP27, and has a strict requirement for the synthesis of viral DNA for expression. The cis-acting regulatory element controlling gC transcription is a 15 bp sequence centered around the TATA box. The requirement for DNA synthesis can be alleviated by fusing the upstream transcriptional regulatory sequences of the early tk gene 5' to the gC TATA element. The requirement for viral DNA synthesis for late gene expression will be the main focus of these experiments. The minimum transcriptional signal capable of giving DNA replication independent expression of the gC gene will be determined. The hypothesis that ICP8, the HSV-1 major DNA binding protein, functions as a general repressor of late gene expression prior to DNA replication will be tested. Using a virus carrying the tk-gC chimeric gene and a temperature sensitive mutant allele of the alpha 27 gene, the role of ICP27 in late gene expression will be probed. Studies have been initiated on the regulation of expression of the gene for the HSV-1 specified large subunit of ribonucleotide reductase (RR1). The RR1 gene shows characteristics of both immediate-early and early gene regulation and, in contrast to all other HSV-1 genes, may be uniquely transactivated by ICP0 instead of ICP4 during infection. The hypothesis will be tested that in infection immediate-early expression is achieved through alphaTIF and the octamer/TAATGARATTC element, while early expression is accomplished through ICP0 and the downstream regulatory signals of the RR1 promoter. The cis-acting signals critical for transactivation by alpha TIF and ICP0 will be defined. Exchange of cis-acting signals between the RR1 promoter and the ICP4 responsive tk promoter should identify the sequences responsive to induction by ICP0 and ICP4. Experiments are proposed to determine if the alpha0 gene product is the only viral transactivator regulating RR1 expression during the early phase of infection. Finally, the RR1 promoter mutant viruses will be tested in mice for effects on neurovirulence and on the establishment and reactivation from latency.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI018228-09
Application #
3127766
Study Section
Experimental Virology Study Section (EVR)
Project Start
1981-08-01
Project End
1994-11-30
Budget Start
1989-12-01
Budget End
1990-11-30
Support Year
9
Fiscal Year
1990
Total Cost
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Type
Schools of Medicine
DUNS #
791277940
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
Makarova, O; Gorneva, G; Wu, F et al. (1996) Incorporation of nuclear matrix attachment regions into the herpes simplex virus type 1 genome does not induce long-term expression of a foreign gene during latency. Gene Ther 3:829-33
Goins, W F; Sternberg, L R; Croen, K D et al. (1994) A novel latency-active promoter is contained within the herpes simplex virus type 1 UL flanking repeats. J Virol 68:2239-52
Gage, P J; Sauer, B; Levine, M et al. (1992) A cell-free recombination system for site-specific integration of multigenic shuttle plasmids into the herpes simplex virus type 1 genome. J Virol 66:5509-15
Dolter, K E; Goins, W F; Levine, M et al. (1992) Genetic analysis of type-specific antigenic determinants of herpes simplex virus glycoprotein C. J Virol 66:4864-73
Levine, M; Krikos, A; Glorioso, J C et al. (1990) Regulation of expression of the glycoprotein genes of herpes simplex virus type 1 (HSV-1). Adv Exp Med Biol 278:151-64
Weber, P C; Levine, M; Glorioso, J C (1990) Recombinogenic properties of herpes simplex virus type 1 DNA sequences resident in simian virus 40 minichromosomes. J Virol 64:300-6
Highlander, S L; Dorney, D J; Gage, P J et al. (1989) Identification of mar mutations in herpes simplex virus type 1 glycoprotein B which alter antigenic structure and function in virus penetration. J Virol 63:730-8
Chrisp, C E; Sunstrum, J C; Averill Jr, D R et al. (1989) Characterization of encephalitis in adult mice induced by intracerebral inoculation of herpes simplex virus type 1 (KOS) and comparison with mutants showing decreased virulence. Lab Invest 60:822-30
Sunstrum, J C; Chrisp, C E; Levine, M et al. (1988) Pathogenicity of glycoprotein C negative mutants of herpes simplex virus type 1 for the mouse central nervous system. Virus Res 11:17-32
Weber, P C; Challberg, M D; Nelson, N J et al. (1988) Inversion events in the HSV-1 genome are directly mediated by the viral DNA replication machinery and lack sequence specificity. Cell 54:369-81

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