Regulation of HPV16 late gene expression during keratinocyte differentiation

角质形成细胞分化过程中 HPV16 晚期基因表达的调控

• 批准号: 7230761
• 负责人: SAMUEL I GUNDERSON
• 金额: $ 16.95万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7230761
• 关键词: 3' Untranslated Regions; Anogenital cancer; Area; Binding; Binding Sites; Biological Assay; Biology; Bovine Papillomavirus-1; Cancer Etiology; Capsid; Cell Line; Cells; Cervix carcinoma; Cessation of life; Codon Nucleotides; DNA; Data; Diagnostic; Diagnostic tests; Disruption; Elements; Environment; Exploratory/Developmental Grant; Gene Expression; Gene Expression Regulation; Gene Proteins; Gene Silencing; Genes; Goals; Heterogeneous Nuclear RNA; Human; Human Papillomavirus; Human papillomavirus 16; Indium; Laboratory Study; Lead; Life Cycle Stages; Link; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Mediating; Messenger RNA; Molecular; Molecular Biology; Nuclear; Outcome; Papillomavirus; Play; Production; Proteins; Publishing; RNA 3' End Processing; RNA Processing; RNA Splicing; Regulation; Regulatory Element; Reporter; Research; Research Personnel; Risk; Role; Site; Small Nuclear Ribonucleoproteins; Staging; System; Tail; Testing; Transfer RNA; Translations; Up-Regulation; Viral; Viral Genome; Virion; Virus; Woman; Work; base; genetic regulatory protein; keratinocyte; keratinocyte differentiation; mRNA Precursor; malignant breast neoplasm; mutant; novel therapeutics; programs; promoter

DESCRIPTION (provided by applicant): Project Summary. The late genes of the papillomaviruses are transcribed throughout all stages of its life cycle. Paradoxically, late gene expression at the mRNA and protein level is restricted to late stages of keratinocyte differentiation when infectious virions are being produced. Although the molecular basis of this restriction is not known, a plausible contributory factor is the Negative Regulatory Element (NRE) found in the viral late gene's 3'UTR. The NREs of Bovine Papillomavirus 1 (BPV1) and human papillomavirus 16 (HVP16) inhibit late gene expression in reporter-based assays in both primary keratinocytes (the natural host cell) or in various cell lines. The BPV1 NRE inhibits by binding to the Ul snRNP splicing factor resulting in inhibition of polyA tail addition. Without a polyA tail, the viral late gene mRNA never matures and is presumably degraded by the nuclear exosome. In contrast to BPV1, the HPV16 NRE is far less understood and is more complicated because it consists of 4 weak, overlapping Ul snRNP binding sites linked to a downstream GU rich region of unknown function. This proposal has two overall goals, first to characterize the HPV16 NRE mechanistically and second to determine its importance to the viral life cycle within the natural environment of differentiating keratinocytes. It is important to note that the regulation of HPV16 genes at the RNA processing level is strikingly understudied when compared to the transcriptional level. The successful completion of this work will rigorously test the importance of the NRE to the viral life cycle and will lead to a mechanistic understanding of NRE function. Relevance. It is estimated that about 400,000 new cases of cervical carcinoma occur worldwide every year, and it is the second leading cause of cancer-related deaths among women after breast cancer. Human papillomavirus (HPV) DNA is found in over 90% of cervical cancers and is also prevalent in anogenital cancers. Epidemiologic and laboratory studies have shown a causative role for HPV and HPV 16 in these cancers. For this reason the molecular biology of the HPV 16 strain has been intensely studied. If this proposal successfully identifies new proteins that regulate viral late gene expression then new therapeutic approaches that disrupt the viral life cycle by disrupting these regulatory proteins can be pursued. Diagnostic tests, based on differential expression of these regulatory proteins, can also be developed.

描述（由申请人提供）：项目摘要。乳头瘤病毒的晚期基因在其生命周期的所有阶段进行转录。矛盾的是，mRNA和蛋白质水平的晚期基因表达仅限于角质形成细胞分化的后期，此时感染性病毒粒子正在产生。尽管这种限制的分子基础尚不清楚，但一个可能的影响因素是在病毒晚期基因的 3'UTR 中发现的负调控元件 (NRE)。在基于报告基因的检测中，牛乳头瘤病毒 1 (BPV1) 和人乳头瘤病毒 16 (HVP16) 的 NRE 可抑制原代角质形成细胞（天然宿主细胞）或各种细胞系中的晚期基因表达。 BPV1 NRE通过结合U1 snRNP剪接因子来抑制，从而抑制polyA尾添加。如果没有多聚腺苷酸尾巴，病毒晚期基因 mRNA 永远不会成熟，并且可能会被核外泌体降解。与BPV1相比，HPV16 NRE知之甚少并且更加复杂，因为它由4个弱的、重叠的U1 snRNP结合位点组成，该结合位点与功能未知的下游富含GU的区域连接。该提案有两个总体目标，首先是从机制上表征 HPV16 NRE，其次是确定其在分化角质形成细胞的自然环境中对病毒生命周期的重要性。值得注意的是，与转录水平相比，HPV16 基因在 RNA 加工水平上的调控研究明显不足。这项工作的成功完成将严格检验 NRE 对病毒生命周期的重要性，并将导致对 NRE 功能的机械理解。关联。据估计，全球每年约有40万例新发宫颈癌病例，是继乳腺癌之后女性癌症相关死亡的第二大原因。人乳头瘤病毒 (HPV) DNA 存在于 90% 以上的宫颈癌中，在肛门生殖器癌症中也很常见。流行病学和实验室研究表明 HPV 和 HPV 16 在这些癌症中具有致病作用。因此，HPV 16 毒株的分子生物学得到了深入研究。如果该提议成功地识别出调节病毒晚期基因表达的新蛋白质，那么就可以寻求通过破坏这些调节蛋白来破坏病毒生命周期的新治疗方法。还可以开发基于这些调节蛋白的差异表达的诊断测试。