Herpes simplex virus type 2 (HSV-2) is a large DNA virus which infects humans and is spread primarily through sexual contact. Cervical carcinoma occurs most frequently in promiscuous women or in women with promiscuous partners. Furthermore, HSV-2 antigens and nucleic acids are detected more frequently in women with cervical cancer than in those without. Thus, HSV-2 may be either the causative agent of cervical cancer or one of several cofactors required to manifest the transformed state. The BamHI-E fragment of HSV-2 induces the neoplastic transformation of established rodent cells and the minimal transforming region of BamHI-E (mtrIII) maps to a 486 base pair fragment. Transformation is mediated by DNA sequence motifs in mtrIII which resemble elements controlling recombination and transcription and not the expression of a viral protein. The questions asked in this proposal are intended to delineate the mechanism of HSV-2 induced transformation. Do cellular proto- oncogenes play a role in HSV-2 mediated transformation? To answer this question, the steady state levels of RNA in transformed cells will be compared to that in normal cells. Do the transcriptional regulatory sequences in mtrIII play a key role in HSV-2 mediated transformation and gene expression? A reporter gene, the bacterial chloramphenicol acetyltransferase gene, will be utilized to correlate the transcriptional enhancer activity of mtrIII with transformation and the expression of HSV-2 genes. Do potential stem-loop structures and DNA conformation in mtrIII mediate neoplastic transformation? Site-specific mutagenesis of a stem-loop structure in mtrIII and the effects of such mutations on transformation will be analyzed. The regions of mtrIII that have altered DNA conformation will be mapped using specific chemical probes and correlated with transformation. Do cells transformed by HSV-2 retain mtrIII DNA sequences? Transformed cells will be analyzed for integrated HSV-2 sequences by Southern blotting experiments and episomal elements which contain HSV-2 DNA will be identified and analyzed in transformed cells. Does HSV-2 posses transforming potential in its natural host, the human? The transforming potential of HSV-2 will be assayed in several human cell lines. In light of evidence which implicates HSV-2 as a causative agent of cervical cancer, these studies will define a structure/function relationship of a novel transforming domain and identify cellular genes that are altered during transformation.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
1R29CA047872-01
Application #
3459047
Study Section
Virology Study Section (VR)
Project Start
1988-07-01
Project End
1993-06-30
Budget Start
1988-07-01
Budget End
1989-06-30
Support Year
1
Fiscal Year
1988
Total Cost
Indirect Cost
Name
University of Mississippi Medical Center
Department
Type
Schools of Medicine
DUNS #
928824473
City
Jackson
State
MS
Country
United States
Zip Code
39216
Hanson, N; Henderson, G; Jones, C (1994) The herpes simplex virus type 2 gene which encodes the large subunit of ribonucleotide reductase has unusual regulatory properties. Virus Res 34:265-80
Garrett, L R; Ackland-Berglund, C E; Jones, C J et al. (1993) Differential effects of TPA and pristane on gene expression and transformation in mouse epidermal cells. Exp Cell Res 205:416-21
Jones, C; Zhu, F; Dhanwada, K R (1993) Analysis of a herpes simplex virus 2 fragment from the open reading frame of the large subunit of ribonucleotide reductase with transcriptional regulatory activity. DNA Cell Biol 12:127-37
Dhanwada, K R; Garrett, L; Smith, P et al. (1993) Characterization of human keratinocytes transformed by high risk human papillomavirus types 16 or 18 and herpes simplex virus type 2. J Gen Virol 74 ( Pt 6):955-63
Bratanich, A C; Jones, C J (1992) Localization of cis-acting sequences in the latency-related promoter of bovine herpesvirus 1 which are regulated by neuronal cell type factors and immediate-early genes. J Virol 66:6099-106
Lee, S H; Ackland-Berglund, C E; Jones, C J (1992) The tumor promoter pristane activates transcription by a cAMP dependent mechanism. Mol Cell Biochem 110:75-81
Garrett, L R; Jones, C J; Cuchens, M A (1992) Pristane induced gene activation. Chem Biol Interact 81:119-30
Dhanwada, K R; Veerisetty, V; Zhu, F et al. (1992) Characterization of primary human fibroblasts transformed by human papilloma virus type 16 and herpes simplex virus type 2 DNA sequences. J Gen Virol 73 ( Pt 4):791-9
Bratanich, A C; Hanson, N D; Jones, C J (1992) The latency-related gene of bovine herpesvirus 1 inhibits the activity of immediate-early transcription unit 1. Virology 191:988-91
Jones, C; Delhon, G; Bratanich, A et al. (1990) Analysis of the transcriptional promoter which regulates the latency-related transcript of bovine herpesvirus 1. J Virol 64:1164-70

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