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: 7414527
• 负责人: SAMUEL I GUNDERSON
• 金额: $ 18.8万
• 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 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7414527
• 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）：项目概述。乳头瘤病毒的晚期基因在其生命周期的所有阶段都有转录。矛盾的是，mRNA和蛋白水平的晚期基因表达仅限于角化细胞分化的晚期，此时传染性病毒粒子正在产生。虽然这种限制的分子基础尚不清楚，但在病毒晚期基因的3'UTR中发现的负调控元件（NRE）可能是一个可能的因素。牛乳头瘤病毒1型（BPV1）和人乳头瘤病毒16型（HVP16）的NREs在基于报告细胞的实验中抑制了原代角化细胞（自然宿主细胞）或各种细胞系的晚期基因表达。BPV1 NRE通过结合Ul snRNP剪接因子抑制polyA尾部添加。如果没有多a尾，病毒晚期基因mRNA就不会成熟，可能会被核外泌体降解。与BPV1相比，HPV16 NRE的了解要少得多，而且更复杂，因为它由4个弱的、重叠的Ul snRNP结合位点组成，这些位点连接到下游一个功能未知的富含GU的区域。该建议有两个总体目标，首先是表征HPV16 NRE的机制特征，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加工水平的调控明显不足。这项工作的成功完成将严格测试NRE对病毒生命周期的重要性，并将导致对NRE功能的机制理解。的相关性。据估计，全世界每年约有40万新发宫颈癌病例，它是仅次于乳腺癌的妇女癌症相关死亡的第二大原因。人类乳头瘤病毒（HPV） DNA在90%以上的宫颈癌中发现，在肛门生殖器癌症中也很普遍。流行病学和实验室研究表明，HPV和HPV 16在这些癌症中起着致病作用。由于这个原因，HPV 16株的分子生物学已经得到了深入的研究。如果这一建议成功地鉴定出调节病毒晚期基因表达的新蛋白，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差异表达的诊断测试。