Abstract

The autonomously replicating parvoviruses contain linear single-stranded DNA genomes of about 5 kilo bases. They are widespread and infect invertebrates, birds, and mammals including man. The parvovirus nonstructural protein NSI has a central role in regulating parvovirus replication. NS1 has been assigned 6 phenotypes: 1. positive regulation of the coat protein promoter, 2. negative regulation of its own gene and heterologous gene expression, 3. requirement for parvovirus DNA replication, 4. requirement for excision if viral termini from a plasmid, 5. inhibition of SV40 DNA replication, and 6. inhibition of cell DNA replication. Our evidence suggests that all or nearly all parvovirus DNA replication requires DNA polymerase delta. We propose to use site-specific mutagenesis to study the roles of NSI in viral DNA replication and its interaction with pol delta. Parvovirus DNA replication is restricted in normal human fibroblasts and this restriction is relieved by neoplastic transformation. The defect in normal cells is at the level of viral DNA replication and it cannot be ascribed to a deficiency in viral] gene expression, rather it appears to result from a deficiency in nuclear translocation of NSI. This project will study the functions of DNA polymerase delta and NSI in viral and cellular DNA replication and explore the defect in viral DNA replication between normal and neoplastically transformed cells.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI025552-05
Application #
2062997
Study Section
Virology Study Section (VR)
Project Start
1990-01-01
Project End
1995-12-31
Budget Start
1994-01-01
Budget End
1995-12-31
Support Year
5
Fiscal Year
1994
Total Cost
Indirect Cost

  Institution

Name
University of Nebraska Medical Center
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
City
Omaha
State
NE
Country
United States
Zip Code
68198

  Publications

Tauer, T J; Schneiderman, M H; Vishwanatha, J K et al. (1996) DNA double-strand break repair functions defend against parvovirus infection. J Virol 70:6446-9
Faisst, S; Faisst, S R; Dupressoir, T et al. (1995) Isolation of a fully infectious variant of parvovirus H-1 supplanting the standard strain in human cells. J Virol 69:4538-43
Dai, Y; Gold, B; Vishwanatha, J K et al. (1994) Mimosine inhibits viral DNA synthesis through ribonucleotide reductase. Virology 205:210-6
Kris, M; Jbilo, O; Bartels, C F et al. (1994) Endogenous butyrylcholinesterase in SV40 transformed cell lines: COS-1, COS-7, MRC-5 SV40, and WI-38 VA13. In Vitro Cell Dev Biol Anim 30A:680-9
Li, X; Rhode 3rd, S L (1993) The parvovirus H-1 NS2 protein affects viral gene expression through sequences in the 3' untranslated region. Virology 194:10-9
Gu, M L; Rhode, S L (1992) Trans-activation of H-1 parvovirus P38 promoter is correlated with increased binding of cellular protein(s) to the trans-activation responsive element (tar). Virology 190:116-23
Gu, M L; Chen, F X; Rhode, S L (1992) Parvovirus H-1 P38 promoter requires the trans-activation region (tar), an SP1 site, and a TATA box for full activity. Virology 187:10-7
Hanson, N D; Rhode 3rd, S L (1991) Parvovirus NS1 stimulates P4 expression by interaction with the terminal repeats and through DNA amplification. J Virol 65:4325-33
Gu, M L; Rhode, S L (1991) Autonomous parvovirus DNA replication requires topoisomerase I and its activity is increased during infection. J Virol 65:1662-5
Li, X; Rhode 3rd, S L (1991) Nonstructural protein NS2 of parvovirus H-1 is required for efficient viral protein synthesis and virus production in rat cells in vivo and in vitro. Virology 184:117-30

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