MECHANISTIC APPROACHES TO HSV-2 INDUCED TRANSFORMATION

HSV-2 诱导转化的机制方法

• 批准号: 3459050
• 负责人: CLINTON J JONES
• 金额: $ 8.66万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-07-01 至 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3459050
• 关键词: Herpes simplex disease; acetyl coA acetyltransferase; binding proteins; cervix neoplasms; chloramphenicol; conformation; disease /disorder model; gene expression; gene mutation; genetic mapping; genetic transcription; herpes simplex virus 2; human tissue; laboratory mouse; mutagens; nucleic acid probes; nucleic acid sequence; oncogenes; oncogenic virus; plasmids; protooncogene; reporter genes; southern blotting; tissue /cell culture; transcription factor; transposon /insertion element; viral carcinogenesis; virus DNA; virus genetics; virus protein; virus replication

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.

单纯疱疹病毒2型（HSV-2）是一种大型DNA病毒